Biochem Combined

Objectives
The student should be able to:
 Describe aerobic and anaerobic glycolysis

 Explain the importance, the fate of the products and important intermediates of
aerobic and anaerobic glycolysis
 Explain the regulation of glycolysis
Dr S Nayak 1
CARBOHYDRATE METABOLISM
Glucose speaks:
“I burn myself to provide fuel to life!
Generated through gluconeogenesis by my friends;
Engaged in the synthesis of lipids, amino acids;
Deranged in my duties due to diabetes mellitus.”
Glycolysis: Degradation of glucose to pyruvate (Lactate under

anaerobic) generates 8 ATP.
Citric acid cycle: The oxidation of acetyl CoA to CO2
Gluconeogenesis: The synthesis of glucose from non-carbohydrate
precursors (amino acids, glycerol etc)
Glycogenesis: The synthesis of glycogen from glucose
Dr S Nayak
2
Glycogenolysis; breakdown of glycogen to glucose and then to
lactate or pyruvate.
Hexose monophosphate shunt: alternative pathway to
glycolysis and TCA cycle for the oxidation of glucose. Here the
glucose is directly oxidised to CO2 and H2O.
Dr S Nayak 3
GLYCOLYSIS (Embden Meyerhof pathway)
• Carbohydrates are the important energy source of the

body.
• Glucose is the preferred source of energy for most of
the body tissues.
• Brain cells derive energy mainly from glucose
• Pyruvate is the end product of aerobic glycolysis
• Lactate is the end product of anaerobic glycolysis
Site: Cytoplasm
Importance of the pathway

• Takes place in all the cells of the body
• Source of energy in erythrocytes
• Anaerobic glycolysis forms the major source of energy
for muscle during exercise
Dr S Nayak 4
• Provide carbon skeletons for the synthesis of non-
essential amino acids.
• Most of the reactions of glycolysis are reversible
• The entry of glucose from ECF to cell (ICF) is under

the control of insulin
• Glycolysis occurrence is the pre-requisite for the

aerobic oxidation of carbohydrates
• Aerobic oxidation takes place in the cells possessing

mitochondria.
• Glycolysis is the major pathway for ATP synthesis in

tissues lacking mitochondria (erythrocytes)
Dr S Nayak 5
Dr S Nayak 6
Dr S Nayak 7
Steps of Glycolysis
1. Irreversible
Glucokinase Hexokinase
a.Present in liver Present in all tissues
b.Phosphorylation of Glu Phosphorylation of hexoses
c.Low affinity for glucose High affinity for substrates
d.Not inhibited by Glucose 6–P Inhibited by glucose 6- P
•Glucose 6-P: Impermeable to the cell membrane.

• Central molecule with a variety of metabolic fates; glycolysis,
glycogenesis, gluconeogenesis and HMP shunt.
2. Reversible reaction
3. PFK is an allosteric inducible key enzyme and the reaction is

irreversible
Dr S Nayak 8
4. Reversible, 6-carbon compound split into two
3 carbon compounds by aldolase
Both are reversibly convertible by an isomerase
enzyme.
Two molecules of glyceraldehyde 3-phospahte
are obtained from a molecule of glucose.
Bromohydroxyacetone-phosphate can inhibit isomerase
5. Reversible,end product contains a high-energy

bond.
Iodoacetate and arsenite non-competitively inhibit
Glyceraldehyde 3-Phosphate dehydrogenase
6. The high energy of 1,3 bisphosphoglycerate is

trapped to synthesise one ATP molecule
Dr S Nayak 9
7. Reversible reaction
8. Reversible reaction, Mg2+ act as activator

Fluoride can irreversibly inhibit enolase with the
removal of Mg2+
9. High energy of Phosphoenolpyruvate is trapped into

ATP by the pyruvate kinase, irreversible reaction.
Ends with pyruvate in the tissues with mitochondria
(aerobic)
10. If anaerobic conditions prevail, the reoxidation of NADH

formed in reaction 5 is by transfer of reducing equivalents
through respiratory chain to oxygen is prevented and get
reoxidised by conversion of pyruvate to lactate by LDH
Tissues that function under hypoxic conditions produce
lactate e.g. Skeletal muscle, smooth muscle and erythrocytes
Dr S Nayak 10
Steps 5 and 10 are coupled
• Glycolysis is the major source of energy in anaerobiosis

• For smooth operation of the pathway NADH is to be
reconverted to NAD+.
• Formation of lactate allows the regeneration of NAD+
NAD+ reused by glyceraldehyde 3-P Dh. Glycolysis
proceeds even in the absence of oxygen to supply ATP.
• Fate of pyruvate depends on the presence or absence
oxygen in the cells.
• The occurrence of un-interrupted glycolysis is very
important in skeletal muscle during strenuous exercise.
• Brain, retina, renal medulla and GI tract derive their
energy from glycolysis.
• Glycolysis in the erythrocytes leads to lactate production
Dr S Nayak
11
Energetics of glycolysis:
Energy consuming steps are 1 and 3
Hexokinase and phosphofructokinase
catalysed reactions = - 2 ATP
Energy yielding steps are: Step 5
Catalysed by Glyceraldehyde –3-PDH
Oxidative phosphorylation:NADH x 2 = 3ATP x 2 = 6 ATP
Substrate level phosphorylation Steps 6 and 9
Catalysed by Phosphoglycerate kinase &
Pyruvate kinase 2ATP x 2 = 4 ATP
Total ATP in aerobic glycolysis =10 ATP – 2 ATP
= 8 ATP/ glucose
Anaerobic glycolysis: 1 and 3 = 2 ATP used
Steps 6 and 9 2 ATP x 2 = 4–2 = 2 ATP
Dr S Nayak 12
Shuttle pathways:
• If the cytosolic NADH uses malate-aspartate shuttle, 3
ATP are produced. If it uses Glycerol phosphate
shuttle produces 2 ATP.
Regulation
• Insulin favours glycolysis by activating key glycolytic
enzymes like glucokinase, phosphofructokinase(PFK)
& pyruvate kinase
• Glucose–6 P, inhibits hexokinase. This enzyme
prevents the accumulation of glucose 6-phosphate.
Dr S Nayak 13
• PFK-1 is an inducible enzyme that increases its synthesis in
response to insulin and decreases in response to glucagon
Pyruvate kinase is an inducible enzyme that increases its synthesis

in response to insulin and decreases in response to glucagon
Dr S Nayak 14
Role of Fructose 2, 6-bisphosphate and regulation of PFK2
F 2,6 BP is the regulatory factor for controlling PFK and then

glycolysis in the liver.
The function of synthesis and degradation of Fructose 2, 6-BP is brought out by

a single enzyme (with two active sites), which is referred to as bi-functional or
Tandem enzyme.
The activity of these enzymes is controlled by covalent modification. cAMP

phosphorylate the tandem enzyme, which results in the inactivation of PFK2
and activation of Fructose 2,6-bisphosphatase
15
Dr S Nayak
Regulation by PFK-2
Low blood glucose: PFK2 inactive and F 2,6 BP active (cAMP dependent
phosphorylation )
High blood glucose: PFK2 active and F 2,6 BP inactive (dephosphrylation)
There will be no stimulation when Fru-2, 6 bisphosphate decreases, with

low glucose the PFK1 remains inactive.
Dr S Nayak 16
Pasteur effect
The inhibition of glycolysis by oxygen
When anaerobic yeast exposed to air, the glucose
utilisation decreases.
In the aerobic condition the levels of glycolytic
intermediates from fructose 1, 6 bisphosphate onwards
decreases while the earlier intermediates accumulate.
The Pasteur effect is due to the inhibition of PFK. Citrate
and ATP inhibition explains the Pasteur effect.
Dr S Nayak 17
Rapoport Leubering Cycle (BPG Shunt)
Dr S Nayak 18
 The kinase reaction is bypassed in the erythrocytes
 No energy is trapped from 2, 3 BPG
 The BPG when combines with Hb, reduces the affinity

of Hb towards oxygen. In the presence of 2, 3 BPG
oxyhemoglobin unload oxygen more easily in tissues
 Therefore the 2, 3 BPG increases in hypoxic condition

15 to 25% of the lactate formed goes through this
pathway
 In hexokinase deficiency phosphorylation does not

takes place further. So 2,3 BPG decreases. Then
affinity to hemoglobin increases
Dr S Nayak 19
Fate of pyruvate
Under aerobic condition pyruvate is transported into
mitochondria via pyruvate transporter
Pyruvate dehydrogenase Complex
Pyruvate Acetyl CoA
NAD+ NADH+ H+
TPP, FAD, Lipoic acid, CoASH
Lack of TPP leads to accumulation of pyruvate
In thiamine deficient alcoholics, pyruvate converted
to lactate and it leads to lactic acidosis
Lactic acidosis is caused by:

Inherited deficiency of PDH
Inability to reoxidise NADH in the electron transport
chain
Excessive NADH production, e.g., ethanol intoxication
Pyruvate dehydrogenase inhibited by arsenic and
mercuric ions Dr S Nayak 20
Ref: Essentials of Biochemistry, Dr S Nayak
Dr S Nayak 21
Integration of Metabolism
Objectives
At the end of this lecture student should be able to :
Explain the roles of the liver, muscle and adipose tissue in the mobilisation,
inter-conversion, consumption and storage of energy substrates
Describe the integration of metabolism during fed and fasting conditions
DR. SHIVANANDA NAYAK 1

Integration of Metabolism
T h e o rga n i s m s p o s s e s s va r i a b l e e n e rg y d e m a n d s ;
h e n c e t h e s u p p l y i s a l s o e q u a l l y va r i a b l e .
T h e c o n s u m e d m e ta b o l i c f u e l m ay b e ox i d i ze d to C O 2
a n d H 2 O o r sto re d to m e e t t h e e n e rg y re q u i re m e n t s
as per the body needs.
AT P s e r v e s a s t h e e n e rg y c u r re n c y o f t h e c e l l .
MARCH 2020
DR. SHIVANANDA NAYAK

Integration of major metabolic pathways of energy
metabolism
Glycolysis:
Degradation of glucose to pyruvate (Lactate under
anaerobic) generates 8 ATP.
Fatty acid oxidation:

Fatty acid (FA) oxidizes to Acetyl CoA.
Energy is trapped in the form of NADH and FADH2
Amino acid degradation:

When amino acids consumed more than the required, are
degraded to meet the fuel demands of the body. The
glucogenic amino acids can serve as the precursor for the
synthesis of glucose via pyruvate or intermediates of TCA
cycle. The ketogenic amino acids form the precursor for
acetyl CoA.
Citric acid cycle:
Acetyl CoA is the common metabolite, produced from different
fuel sources. It enters citric acid cycle and gets oxidized to CO2.
Most of the energy is trapped in the form of NADH and FADH2.
Oxidative phosphorylation:
The NADH and FADH2, produced in different metabolic
pathways, are finally oxidized in the electron transport chain,
which is coupled with oxidative phosphorylation to generate ATP.
Hexose monophosphate shunt:

Concerned with the liberation of NADPH, which is utilized for
biosynthesis of several compounds, including fatty acids and
ribose sugar, which is an essential component of nucleotides.

Gluconeogenesis:
Many non-carbohydrate compounds serve as precursor for
gluconeogenesis.
Glycogen metabolism: Glycogen is the storage form of glucose,

in liver and muscle. Glycogen serves as a fuel reserve to meet
body needs for a brief period.
The metabolic pathways, in general are controlled by four
different mechanisms:
1.The availability of substrates

2.Covalent modification of enzymes
3.Allosteric regulation
4.Regulation of enzyme synthesis

Integration of metabolism during fed state
The various tissues and organs of the body work in a well-coordinated

manner to meet its metabolic demands (usually 2-4 hours after food
consumption)
The insulin is the hormone that is responsible for controlling the
metabolism in various organs of the human body
6
Liver
It is specialized to serve as the body’s central metabolic
clearing house. After a meal, the liver takes up the
carbohydrates, lipids and amino acids, processes them
and routes to other tissues. The major metabolic
functions of liver, in absorptive state are:
1. Carbohydrate metabolism:
Increased Glycolysis, glycogenesis and HMP shunt
Decreased gluconeogenesis
2. Lipid metabolism:
Increased fatty acid and triacylglycerol synthesis
3. Protein metabolism:
Increased degradation of amino acids and protein synthesis.
7
Adipose tissue
It is regarded as the energy storage tissue.
1.Carbohydrate metabolism:
Increases uptake of glucose, glycolysis and HMP shunt
Increased FA and TG synthesis.
Degradation of TG inhibited.
8
Skeletal muscle:
Major metabolic functions of skeletal muscle, in absorptive
state are:
Uptake of glucose is higher and glycogenesis increased.
FA taken up from the circulation.
3. Protein metabolism:
Incorporation of amino acids into proteins is higher.
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Brain
• Glucose is the only source of fuel in an absorptive state.
About 120 g of glucose is utilized per day.
• Free fatty acids cannot cross the blood-brain barrier; hence
their contribution for the supply of energy to the brain is
insignificant.
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Integration of metabolism during fasting
condition
It is a metabolic stress, which imposes certain

metabolic compulsions on the organism.
The hyperglycemic hormones becomes active to
maintain blood glucose and to provide energy to
the human body

The metabolism is reorganized to meet the new
demands of fasting.
Glucose is the fuel of choice for brain and muscle.

During fasting the carbohydrate is not sufficient to
meet the requirements.
 Protein meet the fuel demands of the body

The triacylglycerol (TG) of adipose tissue is the
predominant energy reserve of the body.
Starvation associated with decreased insulin and

increased glucagon.
Liver in starvation
Increased glycogen degradation & then gluconeogenesis
Dietary fuel is unavailable and no liver glycogen remains to
maintain blood glucose.
There is complete dependence upon hepatic
gluconeogenesis, primarily from lactate and alanine.
FA oxidation increased and the TCA cycle cannot cope up
with the excess production of acetyl CoA, so it is diverted to
ketone body formation. The fuel demands of the brain are
met by ketone bodies.
Glucose Production and Utilization in the Fasting State
15
Adipose tissue in starvation
•Glucose uptake and its metabolism reduced
2.Lipid metabolism:
•Degradation of TG increased which leads to
increased release of FA from the adipose tissue,
which serves as fuel for various tissues (brain is
an exception). Glycerol liberated during lipolysis is used
for glucose synthesis by the liver.
•FA and TG synthesis completely stopped here.

16
Skeletal muscle in starvation
Glucose uptake and its metabolism are lowered.
2.Lipid metabolism:
FA and ketone bodies are utilized as fuel by the
muscle. Prolonged fasting adopted to utilize FA.
3.Protein metabolism:
Muscle proteins are degraded and the amino acids are
utilized for glucose synthesis by liver. Protein
breakdown is reduced if the starvation prolonged.
17
Fate of Amino Acids From Muscle Protein Breakdown in Starvation
18
Brain in fasting
In the early phase of starvation, the brain mostly

dependent on glucose, supplied by liver gluconeogenesis.
This, in turn, depends on the amino acids released from
the muscle protein breakdown.
The Ketones bodies play a central role in prolonged

starvation, replacing glucose as the primary fuel for the
brain and signaling a reduction in protein catabolism
Ref: Essentials of Biochemistry
20
Wish you all the best
Dr. Shivananda Nayak

Ketone body metabolism
Professor Shivananda Nayak

The University of the West Indies
Faculty of Medical Sciences
Department of Preclinical Sciences
Biochemistry unit
Objectives
At the end of this topic, you should be able to
understand:
• Conditions in which ketone bodies are formed

• Formation and utilisation of ketone bodies
• Regulation of ketone bodies
Ketone body formation and utilisation
Acetoacetate, -hydroxy butyrate and acetone are collectively called

as ketone bodies.
The process of formation of ketone bodies in the liver is called
ketogenesis.
Blood level is usually less than 2 mg % in well-fed state.
Increased production of ketone bodies is known as ketosis.
High level of ketone bodies in blood are referred to as ketonemia
More ketone bodies in the urine is called as ketonuria.
Lungs mainly eliminate acetone.
The acetyl CoA formed in fatty acid oxidation enters into TCA cycle only
if fat and carbohydrate degradation are appropriately balanced
Conditions in which ketone body formation are
Prolonged starvation:
During starvation the carbohydrate level will be low. So the
stored fat of the adipose tissue break down to free fatty acids.
The free fatty acids formed enter the liver and undergoes -
oxidation to release acetyl CoA which cannot be utilized by the
liver through TCA cycle due to lack of Oxaloacetate.
In starvation TCA cycle is impaired due to the deficiency of

oxaloacetate which is diverted to glucose synthesis
(gluconeogenesis).
Therefore acetyl CoA converted to ketone bodies to meet the

energy needs.
Uncontrolled diabetes mellitus:
Because of the lack of insulin the carbohydrate metabolism is impaired

The adipose tissue fat becomes the main source of energy and its
degradation is generally accelerated.
This results in the excessive production of acetyl CoA, leading to
accumulation of acetyl CoA and its conversion to ketone bodies
Feeding high fat diet:
Excess breakdown of fatty acids in the liver takes place.

Formation of more acetyl CoA.
Once the acetyl CoA formation exceeds more than the requirement of
the liver tissues it is converted to ketone bodies and exported to
muscle, heart and kidney to meet the energy requirement..
So the peripheral tissues switch over to utilize ketone bodies.
Formation of ketone bodies
Site: Liver mitochondria
Two molecules of acetyl CoA Thiolase Acetoacetyl CoA
CoASH Acetyl CoA
HMG CoA synthase
CoASH
β-OH β-methyl glutaryl CoA [HMG CoA]

HMG CoA lyase
Acetoacetate + Acetyl CoA
 -hydroxy butyrate NADH+H+
dehydrogenase NAD+ Spontaneous reaction
 -hydroxy butyrate CO2+ Acetone

The ratio of hydroxy butyrate to acetoacetate depends on the NADH / NAD+
ratio inside mitochondria. Hydroxybutyrate is a honorary to acetoacetate.
Because it is a β- ketoacid, acetoacetate also undergoes a slow, spontaneous
decarboxylation to acetone
Acetoacetate and -hydroxy butyrate are week acids, which slowly
deplete alkali reserves (bicarbonate) of the body and causes metabolic
acidosis. This condition is known as ketoacidosis.
Utilisation of ketone bodies (ketolysis)

The liver cannot utilize ketone bodies because it lacks the enzyme
Thiophorase or CoA transferase which is required for the activation
of ketone bodies
Acetoacetate and -hydroxybutyrate can be used as a source of energy

in peripheral tissues [kidney, muscle].
The -hydroxybutyrate is reconverted to acetoacetate and the
acetoacetate is then reactivated to acetoacetyl CoA.
Acetoacetyl CoA, formed , is cleaved by thiolase to yield two
molecules of acetyl CoA which can be oxidized in the TCA cycle to
H2O and CO2
β-Hydroxybutyrate
NAD + β-Hydroxybutyrate dehydrogenase
NADH +H+
Acetoacetate
Succinyl CoA CoA transferase or Thiophorase
Succinate ATP CoA-SH
Acetoacetyl CoA Acetoacetate
Thiolase Thiokinase
CoASH
2 Acetyl CoA TCA Cycle
During prolonged starvation brain utilize ketone bodies.

Acetoacetate and -hydroxy butyrate serve as an important source of
energy for skeletal muscle, cardiac muscle, renal cortex etc.
Ketone bodies are water soluble

They are easily transported from the liver to various tissues
 During starvation ketone bodies can meet 50-70% energy
needs of brain.
Regulation
 Glucagon stimulate ketogenesis
 Insulin Inhibit ketogenesis
 The increased ratio of glucagon/insulin in diabetes
mellitus promotes ketone body formation.
Ketogenic substances: Fatty Acids, amino acids
Antiketogenic substances; Glucose, glycerol and
glucogenic amino acids (Glycine, Alanine , Serine,
Glutamate etc)
Reference: Essentials of Biochemistry

Dr.S. Nayak March 2020
THE
EXTRACELLULAR
MATRIX
Elastin, Fibrillin, Fibronectin,Laminin and Keratin
Dr. Neetu Mohan

School of Veterinary Sciences,
University of the West Indies,
St. Augustine
ELASTIN
 provides extensibility and
elastic recoil in tissues
 present in large amounts in
 lungs,
 large arterial blood vessels
 some elastic ligaments
 skin, ear cartilage
 other tissues in smaller
amounts
SYNTHESIS OF ELASTIN
DNA (ELN gene)
TROPOELASTIN
• 700 a.a long

mRNA
• Soluble
• contains many hydrophobic a.a’s
• e.g glycine, alanine, valine, proline
Polypeptide :Tropoelastin
• Lysine residues which form crosslinks
Some of the proline residues

hydroxylated to
hydroxyproline; (there are no
hydroxylysines in elastin)
Tropoelastin secreted out of cell
Crosslink formation between lysines

CROSSLINKING OF TROPOELASTIN
 Three out of four lysine residues are oxidatively deaminated to aldehydes by
lysyl oxidase
H H
O O
═
═
N C C N C C
H H CH2
CH2 Lysyl Oxidase
CH2 CH2
O2 NH3 + H2O
CH2 CH2
Oxidative deamination
CH2 C H
═
+ O
NH3
Lysine Allysine
CROSSLINKING OF TROPOELASTIN
 Condensation of three
allysines with an unmodified
lysine to form a cross-link
 cross-links called
desmosines
 highly insoluble and stable

with low turn over rate
EMPHYSEMA
In alveolar • Found in plasma
walls • Produced by liver,
monocytes and alveolar
macrophages
α-AT deficient because:

Neutrophil α-antiproteinase
Several mutations in the α-AT
elastase α-antitrypsin (α-AT)
gene
• One particular mutation –
Intact Degraded purine base substitution
elastin elastin (GAG → AAG); a glutamic
acid substituted by a lysine
Lung tissue cannot regenerate Smoking – causes oxidation
of an important methionine
residue which is necessary for
binding of α-AT to elastase
FIBRILLIN
 A glycoprotein
 Fibrillin types 1 to 4
 Abnormal fibrillin type1 leads to Marfan syndrome
 Mutations in FBN 1 gene (chromosome 15)
 Abnormal fibrillin type 2 leads to Beal’s syndrome
 Mutations in FBN 2 gene (chromosome 5q23)
 Beals syndrome: arachnodactyly; aortic enlargement etc.
FIBRILLIN
 Produced in fibroblasts; secreted into
extracellular matrix
 Crosslinks between Glutamine and
Lysine (transglutaminase)
 Inter and intramolecular disulphide
linkages
 Fibrillin becomes incorporated into
insoluble microfibrils (10-12 nm fibers)
 Provide a scaffold for deposition of
elastin
 Essential for the formation of

 elastic fibers found in connective
tissue
 zonular fibers of the lens
 elastin fibers in the aorta
Lysine
H
O H
O
═
Fibrillin N C C Fibrillin
═
N C C
H (CH2)4 H (CH2)4
NH3+ transglutaminase
N H
NH2 C
═
O + NH3
═
C O
(CH2)2
H
(CH2)2
H
N C C
═
Fibrillin N C C Fibrillin
O
H
═
O
H
Glutamine
MARFAN SYNDROME
 Inherited disorder affecting connective tissue of the
 eyes
 skeletal system - long limbs
 cardiovascular system (dilation of the ascending aorta)
 CAUSES:
 Mutations in the FBN1 gene
1. misfolding of fibrillin-1
2. normal fibrillin-1 protein binds to another protein,
transforming growth factor beta (TGF-β) for cell signaling
Abnormal fibrillin-1 associated with TGF-β affects
vascular smooth muscle development and the
integrity of the extracellular matrix
 Bovine Marfan syndrome closely resembles human

Marfan syndrome
 Similar clinical signs and pathological lesions
FIBRONECTIN
 Adhesive protein ; attaches cells to a
variety of extracellular matrices (except
type IV to which laminin binds instead)
 Approx. 20 different fibronectin chains
 alternative RNA splicing from a single fibronectin
gene
 High molecular weight glycoprotein
 Fibronectin structure
 protein dimer consisting of two nearly
identical polypeptide chains
 Each chain is 60-70nm long and 2-3nm thick
 Linked by a pair of C-terminal disulfide bonds
FIBRONECTIN
 6 domains
 Each domain has specific
binding sites
 for other matrix macromolecules
 Heparan sulfate, collagen (Types I, II
and III), fibrin
 for receptors on the surface of cells

(integrins) – bound to RGD
sequences
 RGD – arg-gly-asp
LAMININ
 Present in type IV matrix (basal laminae) along

with type IV collagen, heparan sulfate,
proteoglycans and entactin (nidogen 1)
 Anchors cell surfaces to the basal lamina
 STRUCTURE:
 Composed of three chains wound together to form a
crucifix shaped structure
 Has multiple domains to bind cell surface receptors,
heparan sulfate, and collagen (like fibronectin)
SUMMARY
 Extracellular matrix (connective tissues)
Importance of the ECM components:
 Structural proteins
• Cell adhesion
 Collagen, elastin, fibrillin
 Adhesive proteins (cell binding to ECM)
• Cell proliferation
 Fibronectin, laminin • Cell migration
 Hydrated Gel forming proteins and Signaling proteins • Apoptosis
 Proteoglycans • Cell-cell communication
 Proteins that facilitates cell binding to the ECM
 Integrins
KERATIN
 Protects epithelial cells from damage
 Keratin filaments present in keratinocytes in the
cornified layer of epidermis
 α-keratins (softer) - mammals
 Skin - keratinocytes
 Hair
 wool
 horns
 nails
 Claws
 β-keratins (harder)
 Nails, scales and claws of reptiles
 Shells of reptiles (tortoise, turtle, terrapin)
 feathers, beaks, claws of birds
 quills of porcupines
ALPHA - KERATIN
 4 segments in α-helical conformation
 Supercoiled (left-handed superhelix)
 Filaments of multiple copies of keratin monomer
 Filaments stabilized by
 Hydrophobic interactions between non-polar side chains of amino acids (Gly and Ala )
 Hydrogen bonding (Ser)
 Disulphide linkages (Cys) – confers insolubility
BETA - KERATIN
 β-keratins (harder)
 β-pleated sheets twisted together
 β-pleated sheets stabilized and hardened by disulfide bridges
Objectives
At the end of this topic you should be able to
• Describe Oxidation of fatty acids

• Discuss how fatty acids are metabolised to provide energy
1
LIPID METABOLISM
OXIDATION OF FATTY ACIDS
Oxidation of fatty acid takes place in mitochondria where the various enzymes for fatty
acid oxidation are present close to the enzymes of the electron transport chain.
Most important theory of the oxidation of fatty acid is the  oxidation of fatty acid.
2
-Oxidation of fatty acid
 Fatty acids are the rich sources of energy

 Energy is released when fatty acid undergoes
 -oxidation
 The  - carbon atom of fatty acid is oxidized
 It is a cyclic process.
 Oxidation of fatty acid occurs at the  carbon
atom resulting in the elimination of two
terminal carbon atoms as acetyl CoA leaving
fatty acyl CoA (2 carbon atom less than the
original fatty acid)
3
 Active form of fatty acid is called as fatty acyl CoA.
 If the starting fatty acid is palmitic acid, which has 16 carbon

atoms, at a time 2 carbon atoms are removed as acetyl CoA,
 7 cycles of -oxidation occurs to convert palmitic acid (16 c)
into 8 acetyl CoA (2c) molecules.
I) Fatty acid is activated to fatty acyl CoA
Acyl CoA
Synthetase or Thiokinase
Fatty acid Fatty acyl CoA
CoASH ATP AMP+PPi
This reaction occurs in cytosol that is outside the mitochondria.

4
 Fatty acyl CoA formed outside the mitochondria cannot cross
the inner mitochondrial membrane.
 Carnitine, a carrier substance carries the acyl group into the
mitochondrial membrane.
Acyl CoA CoASH

Carnitine Acyl carnitine
Carnitine acyl transferase I
Translocase
Carnitine acyl transferase II
Carnitine Acyl Carnitine
Acyl CoA CoASH
5
6
7
1. Once the activated FA enter the mitochondria, flavoprotein
linked acyl CoA dehydrogenase (DH) removes two hydrogen
atoms from the ,  position, forming , - unsaturated fatty
acyl CoA. This contains a double bond at  and  position.
2. Enoyl CoA hydratase adds a molecule of water at the

double bond position of ,- unsaturated fatty acyl CoA
forming  - hydroxy acyl CoA.
3. In the presence NAD+, -hydroxy acyl CoA dehydroegnase

enzyme oxidises -hydroxy acyl CoA to form  -ketoacyl CoA.
4. Thiolase in the presence of CoASH cleaves of -keto acyl CoA

to yield acetyl CoA and fatty acyl CoA having
2-carbon atom less than the original FA. Newly formed acyl
CoA undergoes another 6 more cycles starting from the first
step and is finally degraded into acetyl CoA molecules.
8
Energetics of  oxidation of palmitic acid:
Palmitic acid  8 molecules of acetyl CoA in seven cycles.

Each round of  oxidation generates a molecule of FADH2 (2
ATP) and NADH+H+ (3 ATP).
Total ATPs produced in 7 rounds of oxidation process is 35.

In addition when each acetyl CoA molecule oxidised in TCA cycle,
12 ATPs are generated.
Per cycle of  oxidation

Step I (FADH2) = 2 ATP
Step III (NADH+H+) = 3ATP
7 cycles of  oxidation [5 ATP x 7] = 35 ATP
ATP/acetyl CoA in TCA cycle = 12 ATP

Number of acetyl CoA formed/palmitic acid = 8 x 12 = 96 ATP
9
Total ATP produced = 96 + 35 = 131 ATP
ATP utilised for activation of fatty acid = - 2 ATP
Net ATP produced = 129 ATP
The Standard free energy of palmitate = 2,340 Cal

129 x 7.3 Cal = 940 Cal
The efficiency of energy conservation by FA oxidation=940 x100

2,340
= 40%
• Fatty acids are predominantly oxidised by the process of

-oxidation in mitochondria.
10
Oxidation of odd chain fatty acids
• It is similar to -oxidation with a difference in the final step, a
three – carbon fragment, propionyl CoA is left behind (in place of
2 carbon unit for saturated fatty acids) which is converted to
succinyl CoA
Methyl malonic aciduria

Cause: Vitamin B12 deficiency
Effect: accumulation of
L-methylmalonic acid in
the blood which results in the
excretion of methyl malonic
acid in the urine
11
Regulation of β-oxidation
• The rate limiting step in the β-oxidation is the formation fatty

acyl-carnitine catalyzed by carnitine acyl transferase-1 (CAT–1). It
is an allosteric enzyme and inhibited by malonyl CoA (first
intermediate in the biosynthesis of fatty acid from acetyl CoA
catalyzed by acetyl CoA carboxylase)
Malonyl CoA concentration increases in a well fed state, which

inhibits CAT-1 and results in decreased fatty acid oxidation
In starvation, due to decrease in the activity of Insulin/glucagon

ratio, acetyl CoA carboxylase is inhibited and concentration of
malonyl CoA decreases, releasing the inhibition of CAT-1 and
permitting more acetyl CoA for oxidation
12
13
Peroxisomal fatty acid oxidation
• Peroxisomes are sub-cellular organelles found in all
nucleated cells.
• Peroxisomes are able to conduct oxidation of long chain

fatty acids. Oxidation of very long chain fatty acids (20–26
carbon atoms) begins in peroxisomes by a process similar
to β-oxidation (completed in the mitochondria).
• The action of acyl CoA dehydrogenase differs, it produces

H2O2 rather than FADH2
14
• Catalase located in peroxisomes converts this H2O2 to water and
molecular oxygen. This process is not linked directly to
phosphorylation and the generation of ATP. Once the long chain
fatty acids reduced to octanoyl-CoA (with 8 carbons in its fatty acyl
chain) leave the peroxisomes, transferred to carnitine through
which it enters mitochondria, where they undergo β-oxidation
15
Clinical importance
Clofibrate, A drug used to treat certain types of
hyperlipoproteinemias, stimulates proliferation of peroxisomes and
causes induction of the peroxisomal fatty acid oxidation
Zellweger syndrome
Rare inborn error of peroxisomal oxidation of fatty acid oxidation
Cause: inherited absence of functional peroxisomes in all tissues
The syndrome is caused by defect in the transport of enzymes
into the Peroxisomes, thus long chain fatty acids (with 26-38
carbons) are not oxidized and accumulate in tissues like brain,
kidney and muscle
16
-oxidation:
• -Oxidation of fatty acid can also occur in human
body mainly in liver and brain by removing one
carbon from carboxyl end.
• No activation step
• Hydroxylation occurs at -carbon atom done by
mono-oxygenase system and then oxidised to keto-acid.
• Keto-acid undergoes decarboxylation generates a molecule
of CO2 and a fatty acid.
• Occurs in the endoplasmic reticulum.
• Does not require any CoA and does not release energy.
• Defect in enzyme system leads to Refsum’s disease.
17
Refsum’s Disease
•Is a rare but severe neurological disorder

•Patients with this disease accumulate large quantities of an unusual
fatty acid, Phytanic acid derived from phytol, a constituent of
chlorophyll
•Also present in milk and animal fats
•Phytanic acid cannot undergo - oxidation due to the presence of a
methyl group on carbon-3
•This fatty acid undergo initial -oxidation to remove - carbon and
this is followed by - oxidation
•Refsum,s disease is caused by a defect in the -oxidation due to
the deficiency of the enzyme phytanic acid oxidase.
•So phytanic acid cannot be converted to a compound that can be
degraded by - oxidation.
•Patients should avoid diet containing chlorophyll
18
-Oxidation
 Minor pathway for the oxidation of long chain fatty acid in
microsomes.
 Occurs from both the ends of fatty acid chain.
 Needs hydroxylase enzymes with NADPH and cytochrome
P- 450
 Dicarboxylic acids are produced during this process.
 It is important when -oxidation is defective
 The dicarboxylic acids are excreted in urine causing
dicarboxylic aciduria
 Unsaturated fatty acid can also be activated and transported

across the inner mitochondrial membrane and undergo
-oxidation.
19
Metabolic fate of acetyl CoA:
Glucose, Pyruvate TCA cycle

Fatty acid synthesis
Acetyl CoA Ketone body synthesis
Cholesterol synthesis
Fatty acid oxidation Steroid hormone
Acetyl CoA is produced by aerobic glycolysis of glucose,

Oxidation of fatty acid via -oxidation.
Acetyl CoA is mainly used in citric acid cycle.
20
SIDS
 The sudden infant death syndrome (SIDS) is an unexpected death of

healthy infants usually overnight.
 The reality of SIDS is not known.

 It is now estimated that at least 10% of SIDS is due to deficiency of
medium chain acyl CoA dehydrogenase .
 Glucose is the principal source of energy soon after eating or feeding
babies. After a few hours the glucose level and its utilization decrease
and the rate of fatty acid oxidation must simultaneously increase to meet
the energy needs. The sudden death infants is due to a blockade in
-oxidation caused by a deficiency in medium chain acyl CoA
dehydrogenase
21
Jamaican Vomiting Sickness
Characterized by: Severe hypoglycemia, vomiting, convulsions, coma and
death
Cause: Eating unripe ackee fruit which contains unusual toxic amino acid,
hypoglycin A
This inhibits the enzyme acyl CoA dehydrogenase and thus -oxidation of
FA is blocked, leading to various complications.
Professor S. Nayak March, 2019

22
Objective
At learning this topic you should be able to:
• Describe the structure and function of

lipoproteins
Lipoprotein Structure and Function
Are conjugated proteins, composed of core and

surface
 LP core
 Triglycerides
 Cholesterol esters
 LP surface
 Phospholipids
 Proteins
 Cholesterol
 Lipids are water insoluble
 Present in the blood in the form of lipoproteins
which are water soluble
 They have an outer polar surface, which makes
them water soluble.
Composition and characteristics
Separation by ultracentrifugation
 Four distinct groups based on their density

 Chylomicron (d<0.96),
 Very low density lipoprotein(VLDL, d=0.96-1.006)
 Low density lipoprotein (LDL, d=1.006-1.063)
 High density lipoprotein (HDL, d=1.063-1.21).
Separation by Electrophoresis
 Based on difference in their mobilization in an electric field

Plasma Lipoproteins Classes & Functions
Chylomicrons
 Synthesized in small intestine
(mucosal cells )
 To mobilize dietary lipids
 Transport dietary lipids
 98% lipid, large sized, lowest
density
 Apo B-48
 Receptor binding
 Apo C-II
 Lipoprotein lipase activator
 Apo E
 Remnant receptor binding
Chylomicron Metabolism
 Nascent chylomicron (apo B-48,

apo–A) before they enter
circulation
 Mature chylomicron (+apo C &
apo E)
 Lipoprotein lipase found on the
surface of endothelial cells lining
the capillaries in muscle and
adipose tissues removes the fatty
acids of triglycerides
 Chylomicron remnant
 Apo C removed
 Removed in liver
 Substantial portion of the phospholipid,
apo-A and apo-C are transferred to HDLs
during the process of fatty acid removal
 Chylomicron remnant containing primarily
cholesterol.
 apo-E and apo-B-48 are taken up by the
liver though the interaction with the
chlyomicron remnant receptor
 Very Low Density Lipoprotein (VLDL)

 Synthesized in liver
 Transport endogenous triglycerides (liver to peripheral
tissues
 90% lipid, 10% protein
 Apo B-100
 Apo C-II
 LPL activator
liberates free fatty acids that are taken up by the adipose tissue
and muscle
 Apo E
VLDL Metabolism
 Nascent VLDL (B-100) + HDL (apo C & E) = VLDL

 LPL hydrolyzes TG forming IDL
 IDL loses apo C-II (reduces affinity for LPL)
 75% of IDL removed by liver
 Apo E and Apo B mediated receptors
 25% of IDL converted to LDL by hepatic lipase
 Loses apo E to HDL
 Intermediate Density Lipoprotein (IDL)

 Synthesized from VLDL during VLDL degradation
 Triglyceride transport and precursor to LDL
 Apo B-100
 Apo C-II
 LPL activator
 Apo E
 Low Density Lipoprotein (LDL)

 Synthesized from IDL
 Half life of LDL in blood is 2 days
 transport Cholesterol from liver to
peripheral tissues
 75% of the plasma cholesterol is
incorporated into the LDL particles
are derived from VLDL, a small
part is directly released from liver
 78% lipid (58% cholesterol & CE)
 Apo B-100
Interaction of LDL with LDL receptor
LDL Metabolism
 LDL receptor-mediated endocytosis
 About 75% of LDL are taken up by
the liver, adrenal and adipose
tissue cells by LDL receptor
mediated endocytosis
 LDL receptors on ‘coated pits’
 Clathrin: a protein polymer that
stabilizes pit
 Endocytosis
 Loss of clathrin coating
 uncoupling of receptor, returns to
surface
 Fusing of endosome with lysosome
 Frees cholesterol & amino acids
 High Density Lipoprotein (HDL)

 Synthesized in liver and intestine as
protein rich discoid particles
 Reservoir of apoproteins
 Reverse cholesterol transport
 52% protein, 48% lipid, 35% C & CE
 Apo A
 Activates lecithin-cholesterol
acyltransferase (LCAT)
 Apo C
 Activates LPL
 Apo E
HDL Metabolism: Functions
 Apoprotein exchange
 provides apo C and apo E from VLDL and
chylomicrons
 Reverse cholesterol transport
 Discoid HDLs are converted into spherical
lipoprotein through the accumulation of
cholesterol ester.
Reverse cholesterol transport
 Uptake of cholesterol
from peripheral tissues
(binding by apo-A-I)
 Esterification of HDL-C
by LCAT
 LCAT activated by apoA1
 Transfer of CE to
lipoprotein remnants
(IDL and CR) by CETP
 removal of CE-rich
remnants by liver,
converted to bile acids
and excreted
Hyperlipoproteinemias
Presence of excessive amounts of lipoproteins

like VLDL, LDL, or chylomicrons in the plasma
following a 10 to 12 hours of fasting.
Fredrickson’s Classification:
Type I hyperlipoproteinemia: High TG and Chylomicron
Cause: lipoprotein lipase absence
 Type II a hyperlipoproteinemia: high LDL with high
cholesterol
 Type II b hyperlipoproteinemia: high LDL
(cholesterol) and VLDL (TG)
 Type III hyperlipoproteinemia: high IDL +LDL
 Type IV hyperlipoproteinemia: high-level of VLDL
(high-level of cholesterol and triglyceride)
 Cause: overproduction of endogenous TG.
Diabetes, obesity, chronic alcoholism and renal
failure
 Type V hyperlipoproteinemia: high chylomicrons
and VLDL (high TG).
-obesity
-diabetes mellitus
-alcoholism
-nephrotic syndrome
Familial hypercholesterolemia
 It is a genetic disorder characterized by high

cholesterol levels, specifically very high levels of
low-density lipoprotein, in the blood and early
cardiovascular disease.
 Many patients have mutations in the LDLR gene
that encodes the LDL receptor protein (which
normally removes LDL from the circulation), or
apolipoprotein B (ApoB), which is the part of LDL
that binds with the receptor
 Patients who have one abnormal copy (are
heterozygous) of the LDLR gene may have
premature cardiovascular disease at the age of 30
to 40. Having two abnormal copies (being
homozygous) may cause severe cardiovascular
disease in childhood.
Physical signs
High cholesterol levels normally do not cause any

symptoms. Cholesterol may be deposited in various
places in the body that are visible from the outside,
such as in yellowish patches around the eyelids, the
outer margin of the iris and in the form of lumps in
the tendons of the hands, elbows, knees and feet.
Cardiovascular disease
Deposition of cholesterol in the walls of arteries
leads to atherosclerosis, the underlying cause of
cardiovascular disease.
The most common problem in FH is the development
of coronary artery disease (atherosclerosis of the
coronary arteries) at a much younger age than would
be expected in the general population. This may lead
to angina pectoris (chest pain) or heart attacks.
Pathophysiology
Normally:
LDL cholesterol circulates in the body for 2.5 days
and subsequently binds to the LDL receptor on the
liver cells, undergoes endocytosis, and is digested.
Synthesis of cholesterol by the liver is suppressed in
through HMG-CoA reductase pathway.
In FH, LDL receptor function is reduced or absent
and LDL circulates for an average duration of 4.5
days, resulting in significantly increased level of
LDL cholesterol in the blood with normal levels of
other lipoproteins. In mutations of ApoB, reduced
binding of LDL particles to the receptor causes the
increased level of LDL cholesterol
1/20/20
MDSC1101 – Digestion & Metabolism
Dr. J. Foster
Biochemistry Unit, Dept. Preclinical Sciences
Faculty of Medical Sciences, U.W.I.
What do you think

“bioenergetics” means?
bio = biology
energetics = branch of physics
that studies energy flow
1
1/20/20
What then is the significance of

“bioenergetics” to us?
} Study of how such energy flows, transforms

and harnessed
} Processes by which the body meets energy demands

e.g. digestion & other metabolism
} Transformation and flow of energy within

biological systems, and their environment
} Concerned with the initial and final energy states
of reactants, not the mechanism or kinetics
} i.e. Biochemical thermodynamics
2
1/20/20
} Thermodynamics - laws & principles describing

the flow and interchanges of heat, energy, &
matter in systems
} Concepts applicable to biological systems

• System – part of the universe we are concerned with
• Surroundings – everything else!
} Three types of systems

• Isolated - cannot exchange matter or energy with its
surroundings
• Closed - may exchange energy, but not matter, with the
surroundings
• Open - may exchange matter, energy, or both with the
surroundings
3
1/20/20
Garrett, Grisham. Biochemistry, 2nd ed © 2000
} 1st Law: the total energy of a system (including

surroundings) remains constant
• energy cannot be gained or lost
• it can be transferred from part to part
• It can be converted from one form to another
} 2nd law: total entropy of a system must

increase for a process to occur spontaneously
4
1/20/20
} What determines how energy flows and

whether reactions occur?
Gibbs free energy (G)
} Gibbs free energy

• energy available to reactants/products in rxn
• determines the feasibility of reactions i.e. direction &
extent (predictive)
} Two forms of G used in chemical reactions

• ΔG, the change in G of rxn
• ΔG°, the standard ΔG (reactants/products @ 1mol/L)
} ΔG° useful only under standard conditions
5
1/20/20
} Given a reaction where A ⇆ B, if ΔG

• is negative – rxn is exergonic*, energy is lost from system,
spontaneous from A → B
• is positive – rxn is endergonic*, energy is required by system
from surroundings for rxn to occur
• equals zero – rxn is at equilibrium; no direction favoured
• also ΔG A→B = – ΔG B→A
} Spontaneous reactions move towards equilibrium
* differ from exothermic/endothermic which relate to only heat
6
1/20/20
} ΔG is determined by two factors

• Enthalpy (ΔH) – change in heat of reactants and
products of a rxn (e.g. chemical bonds)
• Entropy (ΔS) – change in randomness/disorder of
reactants & products
} Neither ΔH or ΔS can predict rxn feasibility

alone
°K = 273 + °C
J/mol
ΔG= ΔH – TΔS
•ΔG= ΔH – TΔS
J/mol/K
J/mol
as Δ S increases, ΔG becomes more -ve
7
1/20/20
} ΔG can also be defined for ideal gas reactions using the

following:
ΔG = ΔG° + RT ln [B]/[A]
At constant P (pressure) & T (absolute) – thermal equilibrium
R = gas constant (8.315 J/mol/K)

In = natural logarithm
[B] = concentration of product
[A] = concentration of reactant
} Note that ΔG and ΔG° can have different signs
} Under standard conditions [A]=[B]= 1 mol/L

• ΔG = ΔG° + RT ln [B]/[A]
• ΔG = ΔG° + RT ln 1 (ln 1 = 0)
• ΔG = ΔG°
} ΔG° is predictive only under standard conditions

} ΔG and ΔG° can differ greatly depending on [A],
[B]
8
1/20/20
} At equilibrium (steady-state)
[A] / [B] = constant = Keq
Thus, ΔG = ΔG° + RT ln([B]/[A])
becomes ΔG = ΔG° + RT lnKeq
} At equilibrium ΔG = 0
0 = ΔG° + RT lnKeq
ΔG° = – RT lnKeq
} Biochemical pathway - series of rxns each

with characteristic ΔG
} Thermodynamically for a pathway
• ΔGpathway can be considered additive
• feasibility depends on sum of individual ΔG’s
} As long as the sum of ΔG is –ve the pathway is

feasible
9
1/20/20
Enzymes
They reduce the activation
energy needed for a rxn
} Many biological systems have rxns with +ve

ΔG’s
} How do biological systems overcome +ve ΔG’s?
} Exergonic reactions are usually coupled with
endergonic ones
} Such coupling of rxns involves using a common
intermediate – an energy coupler
10
1/20/20
• The individual half-reactions in aqueous solution:
ATP + H2O « ADP + Pi DGo' = -31 kJ/mol (exo)
Pi + glucose « glucose-6-P + H2O DGo' = +14 kJ/mol (end)
•Hexokinase catalyses this rxn (active site excludes H2O,

promotes coupled over individual rxns)
ATP + glucose « ADP + glucose-6-P DGo'= -17 kJ/mol
• Adenosine triphsphate (ATP) is the coupler for the reaction
11
1/20/20
} The energy currency of cells – universal coupler
} Intermediate in the rank of high-energy

phosphates
} Allows it to accept and donate energy in

numerous rxns
} Terminal phosphate bonds are “high energy” (~)

• release a large amount of energy on hydrolysis
} Phosphate bonds allow

• ATP to release energy for metabolic processes when
hydrolysed to ADP + Pi
• ADP to store energy from catabolic processes as
chemical potential energy in the form of ATP
12
1/20/20
• High activation energy barrier for its bonds
• means very slow hydrolysis of “high energy”

bonds w/o enzyme catalyst
• provide kinetic stability - essential to ATP’s

role (and other compounds with ~ bonds)
} Rapid hydrolysis (due to low barriers) would

hinder ATP’s role in metabolism
} enzymes lower these barriers, coupling the

rxn with other useful ones
} prevents free energy released from ATP

hydrolysis being wasted
13
1/20/20
Harper’s Biochemistry 26th ed, Appleton and Lange, USA
§ Anabolic – complex molecules

from simple ones (endergonic)
§ Catabolic – simple molecules

from complex ones (exergonic)
§ ATP bridges the 2 types of

metabolism
Lehninger, Biochemistry, 4nd edition © 2005
14
1/20/20
} Redox (reduction-oxidation) rxns are inherently

coupled & involve both
• donating e- (oxidation)
• accepting e- (reduction)
} The two halves of a redox rxn are considered
separately:
2+ 2+ 3+ +
Fe + Cu ↔ Fe + Cu
can be rewritten in half-reactions as
1. Fe2+ → Fe3+ + e-
2. Cu2++ e- → Cu+
} Free energy is also transferred when these

electrons move
} The transfer of e- can be measured as reduction
potential (E)
} E - the tendency of a chemical species to acquire
electrons and thereby be reduced
15
1/20/20
} For a rxn, E can be used to calculate G:

ΔG = -nFΔE
likewise, ΔG o = -nFΔE o
n = number of electrons transferred,

F = Faraday’s constant (96,480 J/V/mol),
E = reduction potential
E o = std reduction potential
} +ve E favours a –ve G (forward rxn)

} –ve E favours a +ve G (backward rxn)
} Note: the Eo can also be calculated using:

Ecell = Ecathode - Eanode
} Going back to this reaction:
2+ 2+ 3+ +
Fe + Cu ↔ Fe + Cu
can be rewritten in half-reactions as

1. Fe2+ + e- → Fe3+ cathode
2+ +
2. Cu + e- → Cu anode
The cathode donates e- and the anode accepts
16
1/20/20

2+ 2+ 3+ +
Fe + Cu ↔ Fe + Cu
standard reduction potentials are

1. Fe3+ + e- → Fe2+ Eo = +0.77v
2. Cu2++ e- → Cu+ Eo = +0.16v
Thus Eocell = +0.77-0.16 = +0.61v

2+ 2+ 3+ +
Fe + Cu ↔ Fe + Cu
ΔG o = -nFΔE o
= -(1)(96,480)(0.61)
= -(1)(96,480)(0.61)
= -58,852 J
17
1/20/20
v A simple pathway has 2 steps: A→B → C , where

4
ΔG° for A → B = -25.7 kJ/mol, and Keq is 2.6x10
3
ΔG° for B → C = +14.2 kJ/mol and Keq is 1.2x10
} What is ΔG° for A → C?
ΔG° = (-25.7) + (14.2) = -11.5 kJ/mol
} Write an eqn for ΔG for A → B taking into account the
concentrations of reactant & product.
ΔG= ΔG° + RTlnKeq
v Calculate the equilibrium constant for the hydrolysis of

lactose to glucose & galactose, if the ΔG° is -18.9 KJ/mol
o
at 25 C.
o o
T in K = C + 273; R = 8.315 J/K•mol
o
G = -RTlnKeq
lnKeq = G/-RT
Keq = e(G/-RT)
= e(-18,900/(-8.315x298))
= e(18,900/2478)
= e(7.62)
= 20.74
18
1/20/20
v Calculate the standard free energy change for the

isomerization of glucose 6-phosphate to fructose 6-
o
phosphate, if the equilibrium constant is 0.75 at 27 C.
} Is this a favourable reaction under standard conditions?
} Harper’s Biochemistry, 26th Ed. - Chapter 10

} Lecture will be online within 24 hrs
19
2/17/20
Pentose Phosphate Pathway

MDSC1101 Digestion & Metabolism
Dr. J. Foster
Biochemistry Unit, Dept. Pre-clinical Sciences
Facult yof Medical Sciences
U.W.I., St. Augustine
Lecture objectives
n State
the major products of the pentose
phosphate pathway and discuss the
importance of this pathway, its intermediates,
and by-products.
n Outline
the steps involved in the pentose
phosphate pathway.
1
2/17/20
Introduction to the Pathway
n What is the Pentose Phosphate Pathway (PPP)?

n alternative catabolic pathway for glucose 6-phosphate
n alternative names: phosphogluconate pathway, hexose
monophosphate shunt
n called a “shunt” as it diverts G-6-P from glycolytic pathway
into pentoses which can later be “returned” to glycolysis
n What is the Pentose Phosphate Pathway (PPP)?

n significance - produces ribose 5-phosphate & NADPH
n unlike glycolysis, it does NOT produce ATP
2
2/17/20
n Significance of PPP differs from cell to cell, e.g.

n R-5-P needed for nucleic acid & coenzyme synthesis (NAD, FADH
etc.) in rapidly dividing cells (e.g. mucosa, skin)
n NADPH used for reductive synthesis of fatty acids in tissue e.g.
liver, adipose, mammary glands
n Liver, adrenal glands, gonads also use NADPH for cholesterol &
steroid anabolism

n NADPH also used to create protective reducing atmosphere in
oxygen-exposed tissue (erythrocytes, eye lens, cornea)
metabolism in tissue
e.g. RBCs
O2 H2O2
superoxide . - . hydroxyl
radical O 2 OH free radical
3
2/17/20

n NADPH also used to create protective reducing atmosphere in
oxygen-exposed tissue (erythrocytes, eye lens, cornea)
NADPH + H + G-S-S-G 2H2O
PPP FAD gluthathione Se gluthathione

reductase peroxidase
2H + NADP+ 2G-SH H 2O 2
n The PPP can be partitioned into 2 phases

n Oxidative (irreversible) phase
n Nonoxidative (reversible) phase
n Both phases occur in the cytosol (with glycolysis &

gluconeogenesis)
4
2/17/20
Oxidative Phase
n Results in production of R-5-P, NADPH & CO2

n Involves several enzymes & intermediate products ,
and is summarized as:
G-6-P + 2NADP+ + H2O → R-5-P + CO2 + 2NADPH + 2H +
Oxidative Phase: Reaction 1
n G 6-P oxidized to 6-phosphogluconolactone by glucose 6-

phosphate dehydrogenase (G6PD)
n produces the first molecule of NADPH

n rxn is irreversible, regulatory step of pathway
n G6PD highly specific for NADP+
n G6PD strongly inhibited by fatty acid esters of CoA & NADPH
5
2/17/20
n G 6-P oxidized to 6-phosphogluconolactone by glucose 6-

phosphate dehydrogenase (G6PD)
Adapted from Biochemistry 2nd ed., Garret & Grisham
n 6-phosphogluconolactone hydrolyzed by gluconolactonase ,

opening the ring
n ring hydrolysis is actually spontaneous due to lactone’s unstable

structure
n lactonase significantly speeds up process
n product formed is 6-phospho-D-gluconate (a sugar acid)
6
2/17/20
n 6-phosphogluconolactone hydrolyzed by gluconolactonase ,

opening the ring
n 6-phospho-D-gluconate is oxidatively decarboxylated by 6-

phosphogluconate dehydrogenase in 2 steps
n sugar acid is dehydrogenated to a ketoacid using NADP+

n second molecule of NADPH produced
n ketoacid then decarboxylated to the pentose sugar ribulose 5-
phosphate
7
2/17/20
n 6-phospho-D-gluconate is oxidatively decarboxylated by 6-

phosphogluconate dehydrogenase in 2 steps
Oxidative Phase – Reaction 4
n Finally ribose-5-P ketoisomerase converts ribulose-5-P to

ribose-5-P via an enediol intermediate
8
2/17/20
Special Topic: PPP & Favism
n “Favism” – a condition 1st assoc. with the popular fava beans

n beans are basic food staple of the Mediterranean & Middle East
n in some, after eating beans erythrocytes lyse 24 - 48 hrs later
n free haemoglobin released into the blood (haemolytic anaemia)
n complications include jaundice & kidney failure
n Similar symptoms arise in some individuals upon

n ingestion of antimalarial primaquine & sulfa antibiotics
n exposure to herbicides
9
2/17/20
n Symptoms linked to a genetic deficiency of G6PD

n G6PD catalyses production of NADPH which is vital for
n reductive biosynthesis
n detoxification of reactive oxygen species (ROSes; eg
superoxide radicals, H2O2)
n Compounds such as quinine, primaquine, high doses of

aspirin (and divicine in fava beans) generate ROSes as
metabolic by-products
n RBCs also sensitive to some infections with similar impact
n In G6PD-deficient individuals this becomes an issue

n reduced NADPH production occurs
n excess H2O2 & other ROSes produced in O2-carrying
erythrocytes
n leads to lipid cell membrane breakdown, oxidative damage of
protein & DNA (haemolytic anaemia)
10
2/17/20
n Condition has its advantages

n malarial parasite Plasmodium falciparum very sensitive to levels
of ROSes not harmful to individuals
n G6PD-deficient individuals thus protected against malaria
n condition most prevalent in SE Asia, tropical Africa, parts of
Middle East – areas of high malaria incidence
Nonoxidative Phase
n Some tissues require more NADPH than pentose phosphate
n In these tissues the nonoxidative phase is used to recycle

glucose 6-P from ribose 5-P
n Reversible phase of the PPP
11
2/17/20
Nonoxidative Phase: Reaction 1
n 1st , ribulose 5-phosphate is epimerised to xylulose 5-

phosphate by ribulose 5-phosphate 3-epimerase :
epimerase
Nonoxidative Phase: Rearrangement
n Next, a set of cyclic rearrangement reactions involving the

xylulose 5-phosphate and several carbon sugars occur
n Reactions catalysed by either one of 2 enzymes –

transketolase & transaldolase
n At the end a net of six 5-carbon sugars are converted to

five 6-carbon sugars
12
2/17/20
Oxidative phase PPP Glucose 6-P phosphohexose

isomerase
MORE NADPH!!
1 x 5C 1 x 6C
Xylulose 5-P Glyceraldehyde 3-P Fructose 6-P
TRANSKETOLASE TRANSALDOLASE
Ribulose 5-P
(TPP-dependent)
Ribose 5-P Sedoheptulose 7-P Erythrose 4-P

1 x 5C
TPP = thiamine pyrophosphate
(derived from vitamin B)
2x 5c == 1x 6C
2nd round of non-oxidative

phase in PPP
Glyceraldehyde 3-P
1 x 5C
Glyceraldehyde 3-P Xylulose 5-P
TRANSKETOLASE
(TPP-dependent)
1 x 6C
Fructose 6-P
Fructose 6-P Erythrose 4-P
phosphohexose
isomerase
1 x 6C
MORE NADPH!!
Glucose 6-P Oxidative phase PPP
6x 5c
3x == 4x 6C
5x
2x
13
MDSC 1102
Cardiovascular and Renal
Lecture 1
Nucleotide Metabolism
April 2020 Melford John PhD

Why nucleotide metabolism?
Learning objectives summary
To be able to give an account of:
• the key regulatory points of nucleotide

metabolism;
• metabolic disorders and how they are treated;
• how some drugs work.
Topics
• Review of the 5 bases, RNA, and DNA
• Synthesis of pyrimidine nucleotides
• Synthesis of purine nucleotides
• Degradation of pyrimidine nucleotides
• Degradation of purine nucleotides
• Nucleotide disorders
Nucleotide disorders
Include:
gout;
Lesch-Nyhan syndrome;
orotic aciduria;
severe combined immunodeficiency (SCID).
Review of the 5
bases, RNA and DNA
Pyrimidine and purine
pyrimidine purine
uridine thymine cytosine adenine guanine
RNA RNA RNA RNA

DNA DNA DNA DNA
Pyrimidine nucleosides
CH3
TMP
pyrimidine
ribose-5-P
NH2
ribose-5-P
CMP
UMP
ribose-5-P
Purine nucleotides AMP
NH2
IMP
H
xxx
ribose-5-P
ribose-5-P
xxx GMP
ribose-5-P
purine
NH2
H
xxx
ribose-5-P
ribose-5-P
H
NH2- donated from aspartate
aspartate fumarate
NH2- donated from glutamine
glutamine glutamate
DNA base pairs
thymidine
T
adenosine
A
cytosine
C
guanosine
G
DNA
base pairs
Ribose
used to build DNA used to build RNA

Ribose and PRPP
PRPP
synthetase
ribose-5-phosphate PRPP
Harper’s Biochemistry 26th Ed. © 2003

Naming nucleotides
base
nucleotide adenine
adenosine
monophosphate
AMP
C1
nucleoside
phosphate adenosine
ribose
Naming nucleotides
Nucleoside Nucleotide (NTP)
Molecule Base
(sugar) (phosphoryl)
RNA
purine adenine adenosine adenosine triphosphate
purine guanine guanosine guanosine triphosphate
pyrimidine cytosine cytidine cytidine triphosphate
pyrimidine uracil uridine uridine triphosphate
DNA
purine adenine deoxyadenosine deoxyadenosine triphosphate
purine guanine deoxyguanosine deoxyguanosine triphosphate
pyrimidine cytosine deoxycytidine deoxycytidine triphosphate
pyrimidine thymine thymidine thymidine triphosphate
Synthesis of nucleotides
Two types of pathways are used:
• denovo synthesis – starts with simple metabolic
precursors;
• salvage
pathways - recycles free bases and
nucleosides.
Nucleotide monophosphate
to nucleotide diphosphates
specific NMP kinases
NMP ⇆ NDP
dNMP ⇆ dNDP
Nucleotide diphosphate
to nucleotide triphosphates
nucleotide diphosphate kinase

(NDPK)
NDP ⇆ NTP
dNDP ⇆ dNTP
Formation of deoxy-nucleotides
ribonucleotide
reductase
UDP dUDP
CDP dCDP
ADP dADP
GDP dGDP
ATP, dATP
Ribonucleotide
Reductase
NDP
ATP, dATP, dGTP, dTTP
22
de novo synthesis of
pyrimidine nucleotides
Pyrimidine synthesis
asp gln PRPP carbonate
UMP
uridine nucleotides
thymine nucleotides
cytosine nucleotides
Pyrimidine and purine synthesis
asp gln PRPP carbonate asp gln PRPP gly
UMP IMP
U-nucleotides A-nucleotides
C-nucleotides G-nucleotides
T-nucleotides
mes from and Aspartate
Precursors of pyrimidineresidue
synthesis
glutamine
aspartate
bicarbonate
nitrogenous rings (nitrogenous bases) are cobbl
umber of Precursors
sources: of purine synthesis
glycine
aspartate
formate
formate
glutamine
Synthesis of pyrimidine bases overview
Twelve steps:
steps 1 to 6: precursors -> UMP;
step 7: UMP -> UDP;
—— Branch point ——
step 8 to 9: UDP -> CTP;
steps 10 to 12: UDP -> TMP.
28
Synthesis of pyrimidine UMP to CTP
CO₂ + gln + ATP
1 to 6
CT UMP
Ps
O ynt 7 10
h eta
se UDP dUDP
8 11
9 NH2
UTP dUMP
9
12
CTP TMP
Synthesis of pyrimidine UMP to TMP
O
CO₂ + gln + ATP
1 to 6 ribonucleotide
reductase
UMP
7 10
UDP dUDP
12
8 11
UTP dUMP
thymidylate
9 NH2 CH3
12 synthase
CTP TMP
dUMP to TMP
tetrahydrofolate dihydrofolate
serine reductase
THF
glycine
methylene-THF DHF
dihydrofolate
dUMP TMP -> TDP -> TTP

thymidylate
synthase
Serine provides methyl group
Fluorouracil
thymidylate
synthase
dUMP TMP
fluorodeoxyuridylate
F-dUMP
Methotrexate
dihydrofolate
reductase
DHF THF
Megaloblastic anaemia
• Very large red blood cells
• Inner contents of cells not completely developed
Steps of pyrimidine synthesis
Steps 1 to 4:
formation of orotate ring.
Steps 5:
orotate attached to ribose.
Steps 6:
orotate decarboxylated to form UMP.
Steps 7 onwards:
conversion of UMP to UTP, CTP and TMP.

Steps 1 to 6 O
2 ATP + HCO3- + Glutamine + H2O C

HN CH
2 ADP +
O
Glutamate +
Pi
Carbamoyl
Phosphate 5 C C
Synthetase II
C O N
1 HN CH PRPP PPi 2-
O3P O CH2
COO
O
NH2
C C Orotate Phosphoribosyl H H β
N Transferase
O C O H H
H COO OH OH
O PO3-2 Orotidine-5'-monophosphate
Orotate
(OMP)
Carbamoyl Phosphate
Reduced
Aspartate
Aspartate
4 Quinone
Dihydroorotate
OMP
2
Transcarbamoylase
(ATCase)
Dehydrogenase 6 CO2
Decarboxylase
Pi Quinone O
C
O HN CH
O
C C CH
HO C 3 HN CH2 O N
CH2
NH2 H2O
2-
O3P O CH2
C CH O
H H β
C CH O N
Dihydroorotase
O N H COO H H
H COO OH OH
Dihydroorotate Uridine Monophosphate
Carbamoyl Aspartate
(UMP)
Step 1
-
2 ATP + HCO3 + Glutamine + H2O
2 ADP +
Carbamoyl
carbamoyl
Glutamate +
Phosphate
phosphate
Pi
Synthetase
synthetaseIIII
NH2
O C
O PO3-2
Carbamoyl Phosphate
NH2
carbamoly Phosphate
Step 2
O C
O PO3-2
Carbamoyl Phosphate
aspartate
Aspartate
Aspartate
aspartate
Transcarbamoylase
(ATCase)
transcarbamoylase
Pi
HO C
CH2
NH2
C CH carbamoly Aspartate
O N
H COO
Carbamoyl Aspartate
Steps 3
O
O
C
HO C H2O
HN CH2
CH2
NH2
C CH
C CH Dihydroorotase O N
O N H COO
H COO
Dihydroorotate
Carbamoyl Aspartate
mes from and Aspartate
Precursors of pyrimidineresidue
synthesis
glutamine
aspartate
bicarbonate
O
Step 4 HN
C
CH
orotate
C C
O N
H COO
ReducedOrotate
Quinone
Dihydroorotate
Dehydrogenase
Quinone
O
C
HN CH2
dihydroorotate
C CH
O N
H COO
Step 5 O
C
HN CH
O
C C
C O N
PRPP PPi COO
HN CH 2-
O3P O CH2
O
C C H H β
Orotate Phosphoribosyl
O N Transferase H H
H COO OH OH
Orotate Orotidine-5'-monophosphate
(OMP)
O
Step 6 HN CH
C C
O N
COO
2-
O3P O CH2
O
H H β
H H
OH OH
Orotidine-5'-monophosphate
(OMP)
O
OMP
C
Decarboxylase CH
HN
CO2
C CH
O N
2-
O3P O CH2
UMP H
O
H β
H H
OH OH
Uridine Monophosphate
Case study …
Young male patient presents:
• retarded growth;
• severe anaemia;
• possibly slight mental retardation;
• excessive excretion of orotic acid in urine;
• megaloblastic anaemia not cured by
administration of folic acid.
45
Synthesis of CTP and TMP
UMP
UMP kinase
ribonucleotide reductase
UDP dUDP
nucleoside diphosphate
kinase (NDPK)
UTP dUMP
NH2 methylation thymidylate

CTP synthetase
synthase
CTP TMP
Regulation of synthesis of pyrimidine
-
2 ADP +
Carbamoyl Inhibited by UTP
Glutamate +
Phosphate
Pi Activated by:
Synthetase II
1 • PRPP;
NH2 • ATP
O C
O PO3-2
Carbamoyl Phosphate
NH2
O C Carbamoly Phosphate
ATCase
O PO3-2
Carbamoyl Phosphate
Aspartate
Aspartate
2 Transcarbamoylase Inhibited by CTP
(ATCase)
Pi
Allosterically activated by ATP
O
HO C
CH2
NH2
C CH Carbamoly Aspartate
O N
H COO
Carbamoyl Aspartate
Case study …
• severe anaemia;
49
Orotic Aciduria
• Defect in UMP synthase a protein with activities of:
• orotate phosphoribosyl-transferase;
• orotidylate decarboxylase.
• Type I - deficiency of both activities
• Type II - rarer: only orotidylate decarboxylase

deficiency.
50
Orotic Acidura orotate
O
C
CH
O phosphoribosyl HN
transferase O
C
N
C
C PRPP PPi COO

2-
step 5 HN CH O3P O CH2
H
O
H β
5
Transferase H H
C C OH OH
O N
H COO orotidine Orotidine-5'-monophosphate
monophosphate
(OMP)
orotate
Orotate
6
orotidylate OMP O
Decarboxylase
CO 2
decarboxylase C
HN CH
C CH
O N
step 6 2-
O3P O CH2
O
H H β
UMP H H
OH OH
51
Treatment Of Orotic Aciduria
NH2
1 O C
CO₂ + glutamine + ATP
carbamoyl O PO3-2
phosphate
synthetase II carbamoyl
Carbamoyl Phosphate
phosphate
uridine UMP
inhibits UDP
UTP
CTP
52
Summary
Any Questions
MDSC 1102
Lecture II

de novo Synthesis
of Purine Nucleotides
Purine bases AMP
NH2
xxx
ribose-5P
xxx GMP
IMP NH2
inosine monophosphate xxx
ribose-5P
Pyrimidine and purine synthesis compared
UMP IMP
T-nucleotides
Steps of purine synthesis
Steps 1 to 6:
• creation of smaller ring.
Steps 7 to 10:
• creation of second ring.
Steps 11 plus:
• formation of inosine monophosphate (IMP).
• branch, IMP to AMP and GMP.
O
COO
OOC C
2-
O3P O CH2 H N HC N N
O C4 Aspartate ADP C4
H H α CH + ATP + Pi H
5 CH2 CH
C 5
H OH C
N N
OH OH H2N COO H2N
α-D-Ribose-5-Phosphate (R5P)
Ribose-5-Phosphate
SAICAR Synthetase
8 Ribose-5-Phosphate
ATP Carboxyamidoimidazole Ribotide (CAIR) 5-Aminoimidazole-4-(N-succinylocarboxamide)
Ribose
1 Phosphate
PRPP
Pyrophosphokinase AIR
Car boxylase
ADP + Pi
ribotide (SAICAR)
9
Synthetase
AMP
ATP
+HCO3
7 Fumarate
O
Adenylosuccinate
Lyase
N
2-
O3P O CH2 H HC 4 C
O O H2N N
α O CH
H H C4
5
H O P O P O C CH
OH N C
5
OH
O O
H2N N
H2N
Ribose-5-Phosphate
Ribose-5-Phosphate
5-Aminoimidazole Ribotide (AIR)
5-Aminoimidazole-4-carboxamide
5-Phosphoribosyl-α-pyrophosphate (PRPP)
AIR
ADP + Pi 6 ribotide (AICAR)
Glutamine Synthetase N10-Formyl-

+ H2O
glutamine phosphoribosyl
Amidophosphoribosyl THF
2 Glutamate
Transferase
amidotransferase 2 H
ATP
O THF
AICAR
Transformylase
10
+ PPi N
H 2C CH C
H2N N
2- NH2 C4
O3P O CH2
O CH
H H β C O 5
HN NH C
N
H H O C NH
OH OH H
Ribose-5-Phosphate Ribose-5-Phosphate
β-5-Phosphoribosylamine (PRA)
Formylglycinamidine ribotide (FGAM) 5-Formaminoimidazole-4-carboxamide
Glycine ADP + ribotide (FAICAR)
+ ATP
GAR Synthetase 3 FGAM
Synthetase
Glutamate + Pi
H 2O
IMP
11
ADP
+ Pi
H
ATP +
Glutamine +
H2O
5 O
Cyclohydrolase
H2C NH2 4 H2C

N
CH HN
C
C
4
N
O C CH
HC C5
2-
C O N
O3P O CH2 NH N
O N10-Formyl-THF THF O NH 2-
O3P O CH2
H H O
H H
H H H
OH
GAR Transformylase Ribose-5-Phosphate H
OH OH OH
Glycinamide Ribotide (GAR) Formylglycinamide ribotide (FGAR) Inosine Monophosphate (IMP)

2-
O3P O CH2 H
O
H H α Step 1
H OH
OH OH
ATP
Ribose
PRPP
Phosphate
Pyrophosphokinase
synthetase
AMP
2-
O3P O CH2 H
O O
α O
H H
H O P O P O
OH
PRPP
OH
O O
2-
O3P O CH2 H
O O
α O
H H
H O P O P O PRPP
OH OH
O O
Glutamine
+ H2O Amidophosphoribosyl
Transferase
5-Phosphoribosyl-α-pyrophosphate (PRPP) 2
amidotransferase
Glutamate
+ PPi
2- NH2
O3P O CH2
O
H H β
H
OH OH
H Step 2
2- NH2
O3P O CH2
O
H
H H β
Step 3
H
OH OH
Glycine
+ ATP
GAR Synthetase
ADP
+ Pi H 2C NH2
O C
2-
O3P O CH2 NH
O
H H
H H
OH OH
Glycinamide Ribotide (GAR)

Step 4
H
H 2C NH2 N
H2C CH
O C
N10-Formyl-THF THF
2- C O
O3P O CH2 NH
O O NH
H H
H H GAR Transformylase
OH Ribose-5-Phosphate
OH
Glycinamide Ribotide (GAR) Formylglycinamide ribotide (FGAR)
H
H2C
N
CH Step 5
C O
HN NH
Ribose-5-Phosphate
Formylglycinamidine ribotide (FGAM)
ADP +
Glutamate + Pi
FGAM
Synthetase
H ATP +
N Glutamine +
H 2C CH
H2O
C O
O NH
Ribose-5-Phosphate
Formylglycinamide ribotide (FGAR)

N
HC 4
Step 6
CH
5
C
N
H2N
Ribose-5-Phosphate
ADP + Pi
AIR
H Synthetase
N
ATP
H 2C CH
C O
HN NH
Ribose-5-Phosphate
Formylglycinamidine ribotide (FGAM)
OOC
N
C4
5
CH Step 7
C
N
H2N
Ribose-5-Phosphate
Carboxyamidoimidazole Ribotide (CAIR)
ADP + Pi
AIR
Car boxylase
N
HC 4 ATP
CH +HCO3
5
C
N
H2N
Ribose-5-Phosphate
Step 8
O
COO
OOC
N C
C4 Aspartate ADP HC N N
C4
CH + ATP + Pi
H
CH
C
5 CH2 5
C
N N
H2N COO H2N
Ribose-5-Phosphate SAICAR Synthetase Ribose-5-Phosphate

Carboxyamidoimidazole Ribotide (CAIR) 5-Aminoimidazole-4-(N-succinylocarboxamide)
ribotide (SAICAR)
O
Step 9
COO
C
HC N N
H C4
CH2 CH
5
C
N
COO H2N
Ribose-5-Phosphate
5-Aminoimidazole-4-(N-succinylocarboxamide)
ribotide (SAICAR)
O Fumarate Adenylosuccinate
Lyase
C
H2N N
C4
CH
5
C
N
H2N
Ribose-5-Phosphate
ribotide (AICAR)
O
H2N
C
C4
N Step 10
CH
5
C
N
H2N
Ribose-5-Phosphate
ribotide (AICAR)
N10-Formyl-
O THF
AICAR
C Transformylase
H2N
C4
N THF
CH
5
C
N
O C NH
H
Ribose-5-Phosphate
5-Formaminoimidazole-4-carboxamide
ribotide (FAICAR)
O
H2N
C
C4
N Step 11
CH
5
C
N
O C NH
H
Ribose-5-Phosphate
5-Formaminoimidazole-4-carboxamide
ribotide (FAICAR)
O
H2O
IMP
C Cyclohydrolase
N
HN C
4
CH
5
HC C
N N
2-
O3P O CH2 O
H H
H H
OH OH
Inosine Monophosphate (IMP)

umber of Precursors
sources: of purine synthesis
glycine
aspartate
formate
formate
glutamine
Conversion of IMP to AMP and GMP
GTP GDP
aspartate
IMP fumarate AMP
adenyl succinate synthetase & adenyl succinate lyase
ATP AMP
IMP
dehydrogenase GMP
XMP synthetase
IMP GMP
Salvage of nucleotides
Digestion of DNA
endonucleases
DNA oligonucleotides
phosphodiesterases
oligonucleotides deoxynucleosides
Digestion
of DNA
Nucleosides to free bases
phosphorylases
deoxynucleoside ⇆ base + ribose-1-P

Salvage of free pyrimidine bases
phosphorylase kinase
base ⇆ nucleoside ———> nucleotide
24
Salvage of free bases
Free base Nucleoside eznyme Nucleotide enzyme
uracil uridine phosphorylase uridine kinase
thymine thymine phosphorylase thymidine kinase
inosine HGPRT IMP formed
guanine HGPRT GMP formed
adenine APRT AMP formed

e.g. Salvage of free uracil
uridine phosphorylase
⇆
uracil + ribose-1-P uridine + Pi
⇆
uridine kinase
uridine + ATP UMP + ADP
26
Salvage of purines
hypoxanthine - guanine phosphoribosyltransferase
(HGPRT)
guanine + PRPP ⇆ GMP + PPi
hypoxanthine + PRPP ⇆ IMP + PPi
adenine phosphoribosyltransferase
(APRT)
adenine + PRPP ⇆ AMP + PPi

AMP from free adenine
adenine + PRPP
adenosine
phosphoribosyltransferase
AMP + Pi
28
IMP from free hypoxanthine
GMP from free guanine
hypoxanthine + PRPP guanine + PRPP
hypoxanthine-guanine
IMP phosphoribosyltransferase GMP
(HGPRT)
IMP + Pi GMP + Pi
29
inosine HGPRT IMP formed

Lesch-Nyhan Syndrome
Characterised by:
• neurological and behavioural abnormalities;

• self mutilitation (biting of fingers, toes, and lips);
• overproduction of uric acid that causes:
arthritis;
kidney stones and bladder stones.
31
Deficiency of HGPRT:
• purine bases not salvaged, but broken down;
• produces increased amounts of uric acid.

(HGPRT)
IMP + Pi GMP + Pi
32
Treatment For Lesch-Nyhan
Syndrome
Suggestions?
33
Treatment For Lesch-Nyhan Syndrome
xanthine
xanthine
oxidase
allopurinol
uric acid
34
Nucleotide Degradation
pyrimidine nucleosides purine nucleosides
cytosine GMP AMP

deamination
uracil thymine xanthine
β-alanine β-aminoisobutyrate uric acid

CO2 NH4
Pyrimidine Degradation
cytosine
deamination
uracil thymine
dihydropyrimidine
reduction
dehydrogenase
dihydroxyuracil dihydroxythymine Harper’s Biochemistry 26th Ed. © 2003

Pyrimidine
dihydroxyuracil dihydroxythymine Degradation
hydrolysis
deamination
β-alanine
β-aminoisobutyrate
Purine Degradation
Purine Nucleotide Degradation
• First purine nucleotides lose their phosphates:
AMP → adenosine
GMP → guanosine
• Next:
deamination;
removal of ribose.
Purine nucleoside phosphorylase
purine nucleoside phosphorylases

(PNP)
guanosine guanine + ribose-1-P
inosine hypoxanthine + ribose-1-P

adenosine inosine hypoxanthine
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
PNP xanthine oxidase
guanine xanthine
6
guanine deaminase
uric acid
Gout
• Caused by elevated amounts of urate in blood and tissues
• Leads to abnormal deposition of sodium urate crystals
• Joints become inflamed, painful, and arthritic

• Kidneys especially affected with deposits in tubules
44
Gout
• More common in men than in women:
oestrogen reduces a woman's levels of uric acid
by increasing excretion of uric acid via the
kidneys.
• Estimated 1 in 70 of UK adults have gout:

because incidence of gout increases with age, it
affects 1 in 14 older men and 1 in 35 older
women.
45
What causes gout?
• Abnormalities in renal handling
• Diets rich in red meat and red wine
• Genetic defects in PRPP synthetase also a cause:

• e.g. abnormally high Vmax or feedback-inhibition
resistance;
• defective enzyme causes overproduction of purines.
46
Treatment?
How would you treat the patient?
Any advice on lifestyle changes?

Treatment of Gout
xanthine
xanthine
allopurinol oxidase
uric acid
48
Treatment of Gout
Allopurinol inhibits xanthine oxidase:
• xanthine oxidase converts
allopurinol → oxypurinol
• oxypurinol remains bound to enzyme; allopurinol
• hypoxanthine and xanthine remain:
more soluble than uric acid – less

crystal deposits.
Summary
Any questions?
MDSC 1102
Lecture III

Topics
• Regulation of nucleotide metabolism
• Disorders of pyrimidine metabolism
• Disorders of purine metabolism
• Nucleotide metabolism in relation to cancer
2
Regulation of
Regulation of nucleotide metabolism
Pyrimidine nucleotides Purine nucleotides
• PRPP synthetase • PRPP synthetase
• carbamoyl phosphate • glutamine phosphoribosyl

synthetase II amidotransferase
• aspartate transcarbamoyl • adenylosuccinate synthase

transferase
• IMP dehydrogenase
• CTP synthetase • Ribonucleotide reductase
• Ribonucleotide reductase
Regulation of pyrimidine nucleotides
carbomyl phosphate
synthetase II
CO₂ + glutamine + ATP carbamoyl phosphate
PRPP ATP
ATP
UTP ATCase
CTP
carbamoyl aspartate
UMP ribonucleotide
reductase
UDP dUDP
NDPK
UTP
CTP synthetase CTP
CTP TMP
Regulation of synthesis of PRPP
PRPP
synthetase
ribose 5-phosphate + ATP PRPP
ribose
ADP GDP
Regulation of purine nucleotides
amidotransferase
PRPP + gln phosphoribosylamine + glu
PRPP
AMP GMP
IMP
IMP dehydrogenase
AMP
GMP GMP adenylsuccinate

synthetase
AMP
Glutamine phosphoribosyl
amidotransferase
AMP (or GMP) AMP and GMP
PRPP
active less active inactive

Regulation of purine salvage
IMP phosphoribosyltransferase GMP
(HGPRT)
IMP + Pi GMP + Pi
9
Disorders of pyrimidine metabolism
• Include:
• orotic aciduria (synthesis);
• dihydropyrimidine dehydrogenase deficiency
(degradation).
• Disorders of pyrimidine metabolism are rarer

than those of purine
10
cytosine
Pyrimidine degradation
deamination
uracil thymine
dihydropyrimidine
reduction
dehydrogenase
β-alanine
β-aminoisobutyrate
dihydroxyuracil dihydroxythymine
Dihydropyrimidine dehydrogenase deficiency
Those affected demonstrate:

• neurological problems such as recurrent seizures;
• intellectual disability;
• a small head size (microcephaly);
• increased muscle tone (hypertonia);
• delayed development of motor skills such as walking;
• autistic behaviours.
12
Treatment of cancer
Dihydropyrimidine dehydrogenase deficiency:

• life-threatening toxicity on exposure to 5-FU;
not broken down efficiently;
build up of toxic levels.
• genetic screening before 5-FU treatment.
13
Disorders of purine metabolism
Include:
• Gout
• Lesch–Nyhan syndrome;
• Severe combined immunodeficiency (SCID):
• adenosine deaminase deficiency;
• purine nucleoside phosphorylase deficiency.
14
Severe combined immunodeficiency (SCID)
• Range of mutations that affect immune system
• At least 9 known genes mutations that lead to SCID
• Genetic mutations of enzymes involved in breakdown

and salvage of purine nucleotides cause:
adenosine deaminase (ADA) deficiency;
purine nucleoside phosphorylase (PNP) deficiency.
15
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
guanine xanthine
6
guanine deaminase
uric acid
Adenosine deaminase deficiency
• Also known as bubble boy disease
• David Vetter became famous for living in a sterile
environment in a bubble
17
David Vetter (1971-1984)
18
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
guanine xanthine
6
guanine deaminase
uric acid
ATP, dATP
ADP, GDP, CDP, UDP
Ribonucleotide
reductase
20
ADA deficiency
• A lack of virtually all immune protection
• Neurological problems such as developmental delay,

movement disorders, and hearing loss
• Death at an early age if no treatment that restores

immune function
21
Purine nucleotide phosphorylase (PNP)
• Key enzyme in the:

degradation of guanosine and adenosine;
salvage of guanosine and adenosine.
• Deficiency leads to build up of dGTP that is toxic
to immune system
22
PNP deficiency
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
guanine xanthine
6
guanine deaminase
uric acid
PNP deficiency
• About two-thirds of individuals with PNP deficiency
have neurological problems that may include:
• developmental delay;
• difficulties with balance and coordination (ataxia);
• muscle stiffness (spasticity).
• Also increased risk of autoimmune disorders
24
Drugs
25
Anti-metabolites
• Interfere with DNA and RNA growth
• Target fast-dividing cells to treat:
autoimmune diseases;
cancer.
Methotrexate
dUMP to TMP Trimethroprim
serine reductase
THF
glycine
methylene-THF DHF
dihydrofolatehydrofolate

thymidylate
synthase
5-flurouracil
Drugs
Methrotrexate
folic acid
Drugs
Trimethoprim
Fluorouracil treatment
• Approx 2m patients receive 5-FU treatment worldwide
each year
• Studies show 4-7% Americans exhibit dose-limiting

toxicity that might be associated with a genetic defect
30
Summary
31
Questions
32
MDSC 1102
Lecture IV
Nucleotide Metabolism Revision

Formation of deoxy-nucleotides
ribonucleotide
reductase
UDP dUDP
CDP dCDP
ADP dADP
GDP dGDP
ATP, dATP
Ribonucleotide
Reductase
NDP
3
Learning objectives summary
To be able to give an account of:
• the key regulatory points of nucleotide

metabolism;
• metabolic disorders and how they are treated;
• how some drugs work.
Nucleotide metabolism
• Among earliest pathways
• Five bases conserved over billions of years
• Conserved across 8.7 millions species on planet
• Highly regulated
• Enzymes are targets for drugs
• Susceptible to genetic disorders
Topics
Review:
• synthesis of pyrimidine and purine nucleotides
• degradation of pyrimidine and purine
nucleotides
• nucleotide disorders
5 pyrimidine and purine bases
pyrimidine purine
uridine thymine cytosine adenine guanine
RNA RNA RNA RNA

DNA DNA DNA DNA
Naming nucleotides
base
nucleotide adenine
adenosine
monophosphate
AMP
C1
nucleoside
phosphate adenosine
ribose
Two different pathways:
• denovo synthesis – starts with simple metabolic
precursors;
• salvage
pathways - recycles free bases and
nucleosides.
Pyrimidine Purine
UMP IMP
T-nucleotides
mes from and
Origin Aspartate
of atoms residue
of pyrimidine ring
glutamine
aspartate
bicarbonate
umber ofOrigin
sources:
of atoms of purine ring
glycine
aspartate
formate
formate
glutamine
Synthesis of pyrimidine bases overview
Twelve steps:
steps 1 to 6: precursors -> UMP;
step 7: UMP -> UDP;
—— Branch point ——
step 8 to 9: UDP -> CTP;
steps 10 to 12: UDP -> TMP.
13
Steps 1 to 6 O
2 ATP + HCO3- + Glutamine + H2O C

HN CH
2 ADP +
O
Glutamate +
Pi
Carbamoyl
Phosphate 5 C C
Synthetase II
C O N
1 HN CH PRPP PPi 2-
O3P O CH2
COO
O
NH2
C C Orotate Phosphoribosyl H H β
N Transferase
O C O H H
H COO OH OH
O PO3-2 Orotidine-5'-monophosphate
Orotate
(OMP)
Carbamoyl Phosphate
Reduced
Aspartate
Aspartate
4 Quinone
Dihydroorotate
OMP
2
Transcarbamoylase
(ATCase)
Dehydrogenase 6 CO2
Decarboxylase
Pi Quinone O
C
O HN CH
O
C C CH
HO C 3 HN CH2 O N
CH2
NH2 H2O
2-
O3P O CH2
C CH O
H H β
C CH O N
Dihydroorotase
O N H COO H H
H COO OH OH
Dihydroorotate Uridine Monophosphate
Carbamoyl Aspartate
(UMP)
Step 1 of synthesis of pyrimidine bases
-
2 ADP +
Glutamate +
Carbamoyl
carbamoyl UTP
Phosphate
Pi phosphate
Synthetase II PRPP
synthetase II
ATP
NH2
O C
O PO3-2
carbamoly
Carbamoyl phosphate
Phosphate
Step 2 of synthesis of pyrimidine bases
NH2
carbamoly phosphate
O C
O PO3-2
Aspartate
aspartate
Carbamoyl Phosphate
aspartate
Aspartate CTP
Transcarbamoylase
transcarbamoylase
(ATCase) ATP
Pi (ATCase)
HO C
CH2
NH2
C CH
carbamoly aspartate
O N
H COO
Synthesis of CTP and TMP
UMP
UMP kinase
ribonucleotide reductase
UDP dUDP
nucleoside diphosphate
kinase (NDPK)
UTP dUMP
NH2 methylation thymidylate

CTP synthetase
synthase
CTP TMP
dUMP to TMP
serine reductase
THF
glycine
methylene-THF DHF
dihydrofolate

thymidylate
synthase
Regulation of pyrimidine nucleotides
carbomyl phosphate
synthetase II
CO₂ + glutamine + ATP carbamoyl phosphate
PRPP ATP ATP
UTP ATCase
CTP
carbamoyl aspartate
UMP ribonucleotide
reductase
UDP dUDP
NDPK
UTP
CTP synthetase CTP
CTP TMP
Case study …
• severe anaemia;
20
Orotic Aciduria
• Defect in UMP synthase a protein with activities of:
• orotate phosphoribosyl-transferase;
• orotidylate decarboxylase.
• Type I - deficiency of both activities
• Type II - rarer: only orotidylate decarboxylase

deficiency.
21
Orotic Acidura orotate
O
C
phosphoribosyl HN CH
O
transferase O
C
N
C
C PRPP PPi COO

2-
step 5 HN CH O3P O CH2
H
O
H β
5
Transferase H H
C C OH OH
O N
H COO orotidine Orotidine-5'-monophosphate
monophosphate
(OMP)
orotate
Orotate
6
orotidylate CO
OMP O
Decarboxylase
2
decarboxylase C
CH
HN
C CH
O N
step 6 2-
O3P O CH2
O
H H β
UMP H H
OH OH
22
Treatment Of Orotic Aciduria
NH2
1 O C
CO₂ + glutamine + ATP
carbamoyl O PO3-2
phosphate
synthetase II carbamoyl
Carbamoyl Phosphate
phosphate
uridine UMP
inhibits UDP
UTP
CTP
23
Overview of synthesis of purine bases
Steps 1 to 6:
• creation of smaller ring.
Steps 7 to 10:
• creation of second ring.
Steps 11 plus:
• formation of inosine monophosphate (IMP).
• branch, IMP to AMP and GMP.
2-
O3P O CH2 H
O
H H α Step 1
H OH
OH OH
ATP
Ribose
PRPP
Phosphate
ribose
Pyrophosphokinase ADP GDP
synthetase
AMP
2-
O3P O CH2 H
O O
α O
H H
H O P O P O
OH OH PRPP
O O
2-
O3P O CH2 H
O O
α O
H H
H O P O P O PRPP
OH OH
O O
Glutamine
+ H2O Amidophosphoribosyl
Transferase
5-Phosphoribosyl-α-pyrophosphate (PRPP) 2
amidotransferase
Glutamate
+ PPi
2- NH2
O3P O CH2
O
H H β
H
OH OH
H Step 2
Conversion of IMP to AMP and GMP
GTP GDP
aspartate
IMP fumarate AMP
adenyl succinate synthetase & adenyl succinate lyase
ATP AMP
IMP
dehydrogenase GMP
XMP synthetase
IMP GMP
Regulation of purine nucleotides
amidotransferase
PRPP + gln phosphoribosylamine + glu
PRPP
AMP GMP
IMP
IMP dehydrogenase
AMP
GMP GMP adenylsuccinate
synthetase
AMP
Glutamine phosphoribosyl
amidotransferase
AMP (or GMP) AMP and GMP
PRPP
active less active inactive

Salvage of nucleotides
Digestion of DNA
endonucleases
DNA oligonucleotides
phosphodiesterases
oligonucleotides deoxynucleosides
Salvage of free free bases
phosphorylase kinase
base ⇆ nucleoside ———> nucleotide
32
e.g. Salvage of free uracil
uridine phosphorylase
⇆
uracil + ribose-1-P uridine + Pi
uridine⇆
kinase
uridine + ATP UMP + ADP
33
Salvage of purine bases
hypoxanthine - guanine phosphoribosyltransferase
(HGPRT)
GMP
guanine + PRPP ⇆ GMP + PPi
adenine phosphoribosyltransferase
(APRT)
adenine + PRPP ⇆ AMP + PPi

Characterised by:
• neurological and behavioural abnormalities;

• self mutilitation (biting of fingers, toes, and lips);
• overproduction of uric acid that causes:
arthritis;
kidney stones and bladder stones.
36
Deficiency of HGPRT:
• purine bases not salvaged, but broken down;
• produces increased amounts of uric acid.

(HGPRT)
IMP + Pi GMP + Pi
37
Treatment For Lesch-Nyhan Syndrome
xanthine
xanthine
oxidase
allopurinol
uric acid
38
pyrimidine nucleosides purine nucleosides
cytosine GMP AMP

deamination
uracil thymine xanthine
β-alanine β-aminoisobutyrate uric acid

CO2 NH4
cytosine
deamination
uracil thymine
dihydropyrimidine
reduction
dehydrogenase
dihydroxyuracil dihydroxythymine Harper’s Biochemistry 26th Ed. © 2003

Pyrimidine
dihydroxyuracil dihydroxythymine degradation
hydrolysis
deamination
β-alanine
β-aminoisobutyrate
• First purine nucleotides lose their phosphates:
AMP → adenosine
GMP → guanosine
• Next:
deamination;
removal of ribose.
Purine nucleoside phosphorylase
purine nucleoside phosphorylases

(PNP)
guanosine guanine + ribose-1-P
inosine hypoxanthine + ribose-1-P
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
PNP xanthine
oxidase
guanine xanthine
guanine 6
deaminase
uric acid
Gout
• Caused by elevated amounts of urate in blood and tissues
• Leads to abnormal deposition of sodium urate crystals
• Joints become inflamed, painful, and arthritic

• Kidneys especially affected with deposits in tubules
46
Gout
• More common in men than in women:
oestrogen reduces a woman's levels of uric acid
by increasing excretion of uric acid via the
kidneys.
• Estimated 1 in 70 of UK adults have gout:

because incidence of gout increases with age, it
affects 1 in 14 older men and 1 in 35 older
women.
47
Treatment of Gout
xanthine
xanthine
allopurinol oxidase
uric acid
48
Disorders of pyrimidine metabolism
• Include:
• orotic aciduria (synthesis);
• dihydropyrimidine dehydrogenase deficiency
(degradation).
• Disorders of pyrimidine metabolism are rarer

than those of purine
49
cytosine
deamination
uracil thymine
dihydropyrimidine
reduction
dehydrogenase
β-alanine
β-aminoisobutyrate
dihydroxyuracil dihydroxythymine
Dihydropyrimidine dehydrogenase deficiency
Those affected demonstrate:

• neurological problems such as recurrent seizures;
• a small head size (microcephaly);
• increased muscle tone (hypertonia);
• delayed development of motor skills such as walking;
• autistic behaviours.
51
Treatment of cancer
Dihydropyrimidine dehydrogenase deficiency:

• life-threatening toxicity on exposure to 5-FU;
not broken down efficiently;
build up of toxic levels.
• genetic screening before 5-FU treatment.
52
Disorders of purine metabolism
Include:
• Gout
• Lesch–Nyhan syndrome;
• Severe combined immunodeficiency (SCID):
• adenosine deaminase deficiency;
• purine nucleoside phosphorylase deficiency.
53
Severe combined immunodeficiency (SCID)
• Range of mutations that affect immune system
• At least 9 known genes mutations that lead to SCID
• Genetic mutations of enzymes involved in breakdown

and salvage of purine nucleotides cause:
adenosine deaminase (ADA) deficiency;
purine nucleoside phosphorylase (PNP) deficiency.
54
• Also known as bubble boy disease
• David Vetter became famous for living in a sterile
environment in a bubble
55
ADA deficiency
• A lack of virtually all immune protection
• Neurological problems such as developmental delay,

movement disorders, and hearing loss
• Death at an early age if no treatment that restores

immune function
56
David Vetter (1971-1984)
57
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
PNP xanthine
oxidase
guanine xanthine
guanine 6
deaminase
uric acid
ATP, dATP
ADP, GDP, CDP, UDP
Ribonucleotide
reductase
59
Purine nucleotide phosphorylase (PNP)
• Key enzyme in the:

degradation of guanosine and adenosine;
salvage of guanosine and adenosine.
• Deficiency leads to build up of dGTP that is toxic
to immune system
60
PNP deficiency
2
1
PNP
adenosine
deaminase
3
guanosine
4 5
guanine xanthine
6
guanine deaminase
uric acid
PNP deficiency
• About two-thirds of individuals with PNP deficiency
have neurological problems that may include:
• developmental delay;
• difficulties with balance and coordination (ataxia);
• muscle stiffness (spasticity).
• Also increased risk of autoimmune disorders
62
Drugs
63
Anti-metabolites
• Interfere with DNA and RNA growth
• Target fast-dividing cells to treat:
autoimmune diseases;
cancer.
Methotrexate
dUMP to TMP Trimethroprim
serine reductase
THF
glycine
methylene-THF DHF
dihydrofolatehydrofolate

thymidylate
synthase
5-flurouracil
Drugs
Methrotrexate
folic acid
Drugs
Trimethoprim
Fluorouracil treatment
• Approx 2m patients receive 5-FU treatment worldwide
each year
• Studies show 4-7% Americans exhibit dose-limiting

toxicity that might be associated with a genetic defect
68
Summary
69
Questions
70
Learning objectives
Lecture 1: Macronutrients
Jan 2020 • Describe the components of the diet in terms of
macro and micronutrients
• Explain the term essential nutrients, and outline the

biochemical roles of essential nutrients
• Discuss the concept of recommended daily allowances

(RDA) for energy and essential nutrients, the
derivation of RDA values, and their limitations
• Describe the factors that determine energy and

Melford John PhD protein requirements in man and animals
Biochemistry Unit
Department Of Preclinical Sciences • Describe and compare the major contributors to
University of the West Indies dietary energy intake between man and animals.
1 2
Course assessments and exam

Lectures
• Course Assessment (30 marks):
1. PBL (15 marks);

1. Introduction to nutrition and
macronutrients 2. biochemistry lab (5 marks);
2. Micronutrients: Vitamins 3. spotter exam (10 marks):
3. Micronutrients: Minerals • mainly anatomy and some biochemistry.
• Phase 1A Exam (70 marks):

4. Nutrition related diseases and
assessing nutritional status • 70 MCQs.
3 4
Reading material Definition of food & nutrition
Food is anything that is ingested by an organism

that is used to:
Harper's Illustrated Biochemistry, 30th
Edition (LANGE Basic Science) by Peter support its growth;
J. Kennelly, Robert Murray, Victor provide energy;
Rodwell and David Bender
maintain and repair it.
Nutrition
The sum of the processes by which an organism
utilises food to sustain its existence
5 6
What are nutrients?
Substances obtained from food that are used by the
body:
What are the components of carbohydrates;
a healthy diet? fats and oils;

proteins;
vitamins;
minerals.
Dietary fibres are not nutrients.
7 8
Macronutrients & micronutrients
Nutrients can be divided into two categories

What happens if you do not 1. Macronutrients are required in large quantities:
eat a healthy diet?

carbohydrates, proteins, fats, and oils;
10 - 100 grams per day.
2. Micronutrients are required in minute quantities:
vitamins and minerals;
micrograms - milligrams per day.
9 10
Variety of nutrients Daily energy requirements

All nutrients are required • An average man needs around 2,500 calories a
A nutrient may serve more than one function day to maintain his weight
No single food has all the nutrients
• An average woman, needs around 2,000
calories a day
• These values vary depending on age and levels

of physical activity, among other factors
11 12
Daily energy requirements Energy source
personal energy requirement = basic energy requirement (BER)
+ extra energy requirement (EER)
BER = 1.3 calories x hours x kg body weight

Carbohydrates: 57%
EER = 8.5 calories x hours x kg body weight
Fats: 30%
For a 60 kg person (132 lbs) who exercises for 2 hr every day
Proteins: 13%
= (1.3 x 24 x 60) + (8.5 x 2 x 60)
= 1872 + 1020 = 2892 calories
13 14
Carbohydrates Functions of carbohydrates

Functions of carbohydrates Main function is to provide energy:
Forms of carbohydrates: 40 - 80 % of the total energy intake;
simple carbohydrates; ideal contribution is around 60 %;
complex carbohydrates: 1g provides 4 kcal.
• starch; Other functions:
• glycemic index; glycoproteins;
• which starches to eat; DNA and RNA.
• glycogen.
Sources of carbohydrates
15 16
Forms of carbohydrates Sources of carbohydrates

Carbohydrates are present in two forms: Milk contains lactose, but carbohydrates obtained mostly
from plants
• simple carbohydrates: e.g. glucose, fructose,
sucrose; Rich sources of compound carbohydrates:
• compound carbohydrates: e.g. starch, glycogen. grains (rice, wheat, maize);
Compound carbohydrates are polymers. roots and tubers (potato, sweet potato, guam, banana);
legumes (pulses, nuts).
17 18
Sucrose Fructose
• Absorbed almost entirely by the liver:
- most cells lack its transporter glut-5;
• Does not stimulate insulin release;
• Poorly absorbed from gastrointestinal tract:
- peripheral blood concentration ≈0.01 mmol/L;
- glucose 5.5 mmol/L.
• Does not suppress grehlin;
• Increases plasma TG -> increase in VLDL:
- bad cholesterol build up on the walls of arteries.
19 20
Control of hunger hexokinase
e Fructose
phosphoglucose k inas
isomerase hexo
fructokinase
decreases phosphofructose Fructose 1-phosphate

apetite kinase (PFK)
increases aldolase aldolase

apetite
triode phosphate
isomerase
21 22
Lactose Lactose intolerance

Missing lactase in small intestine
Disaccharide of galactose and glucose:
Fermentation in the gut causes:
• found in milk;
• hydrolysed by lactase. • bloating;
• stomach pains or cramps;
• diarrhoea;
• wind;
• bowel sounds that you can hear;
• feeling sick;
• an urgent need to have a bowel movement.
23 24
Maltose Digestion of disaccharides
• lactose (gal-glu) hydrolysed by lactase
• maltose (glu 1-4 glu) hydrolysed by maltase

Disaccharide of glucose and glucose:
• isomaltose (glu 1-6 glu) hydrolysed by
• alpha 1-4 glycosidic bond; isomaltase
• formed primarily from the partial hydrolysis • sucrose (glu-fru) hydrolysed by sucrase.
of starch;
• found in malt beverages such as beer.
25 26
Amylose
Starch
Linear polymer of glucose

Produced by green plants as an energy store
Makes up 15 to 20 % of starch
Large polymer of glucose with glycosidic bonds
Composed of a mixture of two substances: Consist of 200 to 20,000 residues
• amylose - essentially a linear polysaccharide; Hydrolysed by amylase

• amylopectin - a highly branched polysaccharide. End products are glucose and maltose.
27 28
Which starches to eat?

Amylopectin
Foods that are digested slowly such as:
• grains that are whole or intact when cooked;
• whole beans;
• lentils.
Canned food digested more readily
• Highly branched polymer of glucose
• Side chains of 30 glucose units attached via 1α→6

bonds every 20 to 30 glucose units along chain mung dal
29 30
Which starches to eat? Glycogen
• The equivalent of starch in animals
Eat whole wheat brown breads that have more fibre
Avoid anything with white flour as much as you can • Similar in structure to amylopectin, but more branched
Eat cereal with extra fibre such as bran flakes: • Various samples of glycogen have been measured at
1,700-600,000 units of glucose.
• avoid processed cereals with little fibre.
31 32
Glycogen Glycemic index

• Some glucose not used by the body stored as glycogen in
muscle and liver A measure of the potency of a test food at raising
blood glucose level compared to potency of glucose
• Extra glucose stored as fat
• The body's glycogen capacity is limited to about 350 grams:
• lasts approximately 10 to 12 hours when at rest.
• Glycogen important as under normal conditions glucose only

fuel used by brain.
33 34
Glycemic index
Example of calculation of GI for white bread:
Glycemic index
subject fed 50g of glucose (i.e. 50g carbohydrate)
• GI of 100 common foods
blood glu level after 2 hrs is 180 mg/dl
subject fed 71g of bread (i.e. 50g carbohydrate)
blood glu level after 2 hrs is 126 mg/dl
GI = 100 x (126 / 180) = 70
35 36
Blood glucose
Glycemic load
• Takes size of portions into account
• Indicates how much a given serving of a test food

will raise blood glucose:
GL = (GI / 100) x g of carbohydrate in serving
37 38
GL of watermelon Glycemic load

Diets with low GLs linked to lower risk of heart disease
• GI of watermelon is 72
• Given 120g of watermelon has 6g of

carbohydrate:
GL of this = (72/100) x 6 = 4.32
• Thus 120g watermelon has a low glycemic

load
39 40
Proteins
Functions of proteins
Provide 4 kcal of energy per gram
Building material of body parts such as:
Amino acids not manufactured by the body are termed
• muscle, brain, blood, skin, hair, nails, bones and body
essential amino acids: fluids.
• must be obtained from our diets; Essential for functions such as:
• arginine (required for the young, but not for adults), • growth;
histidine, isoleucine, leucine, lysine, methionine, • repair of worn-out tissues;
phenylalanine, threonine, tryptophan, and valine. • replacement of used-up blood;
• resistance against infections.
41 42
Sources of proteins Fats and oils
Dietary proteins obtained from both animal and plant foods
Rich sources are:

• Triglycerides
• meat, fish, egg, milk, and milk products; • Oils
• pulses, nuts, and beans are rich sources of plant proteins. • Essential fatty acids
Cereals and fruits are low in protein: • Cholesterol
• rice: 6 – 8%
• wheat: 12 – 14%.
43 44
Fats and oils Fats and Oils

• Fats provide the building materials for some body parts,
Concentrated sources of energy: such as brain, cell membranes, and hormones
• 1 gm of fat provides 9 kcal of energy.
• Fats also facilitate absorption, transport and storage of
The term fat includes: fat-soluble vitamins A, D, E and K
• triglycerides; • Like all other nutrients, fats are beneficial if consumed

• phospholipids; in the right amount and if of the right type
• sterols such as cholesterol.
45 46
Trans fatty acids Essential fatty acids

The only fatty acids known to be essential for complete
nutrition in many species including humans
• Trans fats are rare in nature, but can occur in
processes food
• Trans fats increase the risk of coronary heart disease:
• they raise levels of LDL cholesterol;
• they lower levels of HDL cholesterol.
• Trans fats from partially hydrogenated oils are more

harmful than naturally occurring oils.
18 carbon atoms
47 48
Essential fatty acids Essential fatty acids
• Double bonds can be introduced at the Δ4, Δ5, Δ6, and Food sources include:
Δ9 positions in most animals, but not beyond the Δ9
position • oily fish, flaxseed, dark green leafy vegetables,
walnuts, and seeds.
• Plants can introduce double bonds at Δ12 and Δ15.
Oily fish include sardines, anchovies, kippers, mackerel,
and herring
49 50
Dietary fibres Dietary fibres

• Dietary fibres are non-digestible, non-absorbable
components of food • Fruits, vegetables, pulses, and whole cereals are sources
of dietary fibre
• Fibres form the bulk of the stool and help in clearing
the bowel and in preventing constipation and colon • Our daily diet should contain some fibre for good health
cancer and well being
• Fibres slow absorption of glucose and cholesterol from
the GI tract, thus are helpful in diabetes and heart
disease
51 52
Summary
Macronutrients
That’s it?
The problem with fructose
Which type of starch
Essential fatty acids
Trans fatty acids
53 54
Lecture 2: Micronutrients - Vitamins Micronutrients
Jan 2020
Micronutrients are nutrients that are required by

the body in minute quantities:
• micrograms to milligrams;
• consist of vitamins and minerals.
Melford John PhD

Biochemistry Unit
Department Of Preclinical Sciences
University of the West Indies
Fat-soluble and water-soluble vitamins

What are vitamins?
Water-soluble vitamins are:
Vitamins are complex, essential organic substances that: • the 8 B-complex vitamins that help to
• perform a variety of metabolic functions; metabolise carbohydrates, fats and proteins:
• perform the same functions in different forms of life; • vitamin C;

Fat-soluble vitamins are:
• are constituents of food;
• vitamins A, D, E, and K.
• are required in small amounts.
The B-complex vitamins The other vitamins
vitamin C
pantothenic acid (B5) vitamin A
niacin (B3)
thiamine (B1)
riboflavin (B2) vitamin K
vitamin D
pyridoxine (B6)
biotin (B7) folic acid (B9)
cobalamin (B12)
vitamin E
EU Recommended daily allowance
Storage of vitamins
• Excess water-soluble vitamins are excreted in urine
• Excess fat-soluble vitamins are stored in the body
Cooking vitamins
A Danish study on cooking broccoli showed:

B Vitamins
• boiling for 5 minutes caused 45 to 64 % loss of vitamin C;
• steaming for 5 minutes retained almost 100 % water-

soluble vitamins;
• microwaving and stir-frying reduce vitamin loss because

they cook food quickly;
• avoid deep-frying, high heat destroys heat-sensitive vitamins.
Vitamin B name Deficiency Coenzyme
thiamine B1 beriberi TPP
arabiniflavanosis
riboflavin B2 FMN FAD
glossitis
niacin B3 pellagra NAD NADP
pantothenic acid B5 rare CoA
pyridoxine B6 cheilosis glossitis PLP
biotin B7 rare, hair loss
megaloblastic
folic acid B9 THF
anemia
cobalamin B12 pernicious anemia

1.4 mg
Thiamine (B1) Pyruvate dehydrogenase
Coenzyme form of thiamine is thiamine
complex
pyrophosphate (TPP)
Thiamine (B1) 1.4 mg

Thymine pyrophosphate (TPP)
Required as a coenzyme for:
thiamine pyrophosphate
• the α-ketoglutarate dehydrogenase complex (TCA
cycle)
Thymine pyrophosphate (TPP)

1.4 mg
Thiamine (B1) deficiency

Also required for transketolase reactions:
• No significant stores of thiamine in the body
• Deficiency can develop within 2 weeks

Thiamine (B1) deficiency
1.4 mg
Symptoms of dry beriberi
Causes:
• Difficulty walking
• alcoholism and malnutrition; • Loss of feeling (sensation) in hands and feet
• excess alcohol makes it harder to absorb thiamine. • Loss of muscle function or paralysis of the lower legs
Deficiency causes beriberi of which there are 4 types: • Mental confusion/speech difficulties
• dry beriberi (peripheral neuritis); • Pain
• wet beriberi (the heart and circulatory system); • Strange eye movements (nystagmus)
• cerebral beriberi (Wernicke-Korsakoff’s syndrome); • Tingling
• infantile beriberi. • Vomiting
Riboflavin (B2)
1.6 mg
Symptoms of wet beriberi
• Awakening at night short of breath
• Increased heart rate
• Shortness of breath with activity
• Swelling of the lower legs

FMN
riboflavin flavin mononucleotide FAD
flavin adenine dinucleotide
Riboflavin (B2) Riboflavin (B2) deficiency

1.6 mg 1.6 mg
FMN is required for: ariboflavinosis

• l-amino acid oxidase;
glossitis
• cytochrome C reductase.
FAD is required as a coenzyme for:
• pyruvate dehydrogenase complex;
• succinate dehydrogenase;
• α-ketoglutarate dehydrogenase complex;
• xanthine oxidase.
Riboflavin (B2) deficiency
1.6 mg 18 mg
Niacin (B3)
Also characterised by:
• seborrheic dermatitis; Metabolic function is to form the nicotinamide
ring of the coenzymes:
• corneal vascularisation.
• NAD;
• NADP.
Niacin (B3)
18 mg 18 mg
Niacin (B3)
Helps release energy from nutrients
NAD is required as a coenzyme for:
• pyruvate dehydrogenase complex;
• α-ketoglutarate dehydrogenase complex.
NADP is required for reactions involving:
• glucose-6-phosphate dehydrogenase;
• 6-phosphate gluconate dehydrogenase.
B3 niacin
Niacin (B3) deficiency

18 mg 6 mg
Pantothenic acid (B5)
pellagra
6 mg 6 mg
CoA required for reactions that synthesise: Coenzyme A required for:
• essential fats and cholesterol; • pyruvate dehydrogenase complex;
• α-ketoglutarate DH complex;
• steroid hormones;
• thiokinase;
• the hormone melatonin.
• thiolase.
B vitamins 6 mg
Thiamine (B1)
Also required for:

• synthesis of neurotransmitter acetylcholine;
• synthesis of heme component of hemoglobin;
• metabolism of a number of drugs and toxins by
Niacin (B3)
the liver.
Pantothenic Acid (B5)

Riboflavin (B2)
Pyridoxine (B6)
6 mg
Pantothenic acid (B5) deficiency
Helps to make red blood cells
Deficiency in humans is very rare and has only been
Helps in amino acid and fatty acid metabolism:
observed in cases of severe malnutrition
• coenzyme form is pyridoxal phosphate (PLP);
• PLP is required as coenzyme for enzymes such as:
transaminases;
decarboxylases.
Biotin (B7)
0.15 mg
Pyridoxine (B6) deficiency
• Biotin functions as a coenzyme in reactions

involving CO2
• Helps release energy from carbohydrates:
pyruvate carboxylase;
propionyl CoA carboxylase;
acetyl CoA carboxylase.
• Aids fat synthesis
cheilosis glossitis
0.15 mg 0.15 mg
Deficiency of biotin (B7) Deficiency of biotin (B7)
• Deficiency is very rare
• Some initial symptoms are: Important to treat right away because it can
hair loss, dry skin, brittle hair, fungal infections, cause severe problems such as:
seborrheic dermatitis, and rashes.
• changes in mental status;
• Other symptoms include:
• hyperesthesias (abnormal increase in stimuli);
fatigue, anemia, nausea, appetite loss, conjunctivitis,
depression, dandruff, psoriasis, eczema, and loss of • parenthesia (sensation of tingling skin).
muscular reflexes.
Folic acid (B9)

0.2 mg 0.2 mg
Helps in the formation of DNA and new blood cells
Important for growth of foetus
Deficiency causes:
megaloblastic anemia:
• impairment of the methionine synthase reaction leads

to impairment of purine ring synthesis;
• the impaired synthesis of DNA prevents cell division

and formation of the nucleus of new red blood cells.
megaloblastic
growth failure. anaemia
1 µg
Cobalamin (B12)
0.2 mg
Folic acid (B9)
Coenzyme form is tetrahydrofolate (THF): B12 used as descriptor for the cobalamins:
• carrier of single carbon;
• contain cobalt and corrin ring;
• involved in single carbon transfer reactions.
• posses biological vitamin activity.
Some corrinoids have no vitamin B12 activity.
Co
corrin ring
Active forms of cobalamin (B12) Cobalamin (B12) deficiency

Pernicious anaemia arises when cobalamin
methylcobalamin deficiency impairs the metabolism of folic acid:
deoxyadenosylcobalamin • leads to folate deficiency that disturbs

erythropoiesis;
• immature precursors of erythrocytes released

into circulation (megaloblastic anemia).
Pernicious anemia Symptoms of cobalamin (B12) deficiency

The most common cause of pernicious anemia is failure of Symptoms include:
absorption of cobalamin: • diarrhoea or constipation;
• to absorb B12 the body uses a special protein called • fatigue, lack of energy, or light-headedness when
standing up or with exertion;
intrinsic factor, which is released by cells in the
stomach; • loss of appetite;
• B12 bound to intrinsic factor is absorbed in the last part • pale skin;
of the small intestine;
• problems concentrating;
• when the stomach does not make enough intrinsic factor, • shortness of breath, mostly during exercise;
the intestine cannot properly absorb vitamin B12.
• swollen, red tongue or bleeding gums.
Cobalamin (B12) deficiency nerve damage Enzymes dependent on cobalamin (B12)
Nerve damage caused by cobalamin deficiency: Three enzymes dependent on cobalamin:
• methylcobalamin required in conversion of homocysteine • methylmalonyl CoA mutase:

to methionine;
converts methyl malonyl CoA into succinyl CoA;
• causes lack of methionine used to make myelin.
B12 deficiency causes accumulation of methylmalonyl
CoA and urinary excretion of methylmalonic acid, which
Occurs over a long period of time (decades) and may cause: provides a means of assessing vitamin B12 nutritional
status.
• confusion or change in mental status (dementia) in severe
or advanced cases;
• leucine aminomutase
• depression; • methionine synthase:
• loss of balance, numbness and tingling of hands and feet. synthesis of methionine (affects myelin production).
Sources of Cobalamin (B12)

Vitamin B name Deficiency Coenzyme
Sources: thiamine B1 beriberi TPP
• liver, meat, fish, eggs and milk; riboflavin B2

arabiniflavanosis
FMN FAD
glossitis
• human beings get small amounts of vitamin B12
from their intestinal flora. niacin B3 pellagra NAD NADP
pantothenic acid B5 rare CoA
pyridoxine B6 cheilosis glossitis PLP
biotin B7 rare, hair loss
megaloblastic
folic acid B9 THF
anemia
cobalamin B12 pernicious anemia
Vitamin A
• Also known as retinol
Other vitamins • Retinol used to synthesise retinal, without which we

would not be able to see
Retinal synthesised from β-carotene Atomic motion of retinal
β-carotene dioxygenase
dehydrogenase trans-retinal
cis-retinal
• The light energy of a photon is converted into motion

• Six double bonds give retinal property to absorb light
Voet D, Voet JG and Pratt CW.2013. Fundamentals of Biochemistry-Life at the molecular level John Wiley and Sons, Inc., USA
Vitamins A deficiency
Vitamin A functions
Deficiency causes night blindness; complete
Vitamin A is also an antioxidant: blindness can also occur:
• protects against free radicals that attack structural • globally 250,000-500,000 children become
blind each year;
proteins, enzymes, and cell membranes of the lens.
• highest prevalence in Southeast Asia and
Vitamin A maintains mucus membranes lining the eye Africa;
• these membranes provide resistance to bacterial • half of these blind children die within a year.
invasion.
Vitamins A deficiency Vitamins A deficiency

Keratomalacia
Follicular hyperkeratosis:
• keratin deposits form around

hair follicles;
• causes rough texture to the

skin resembling 'goose
flesh;'
• facial skin scaly, off-colour,

cracked, or dry to the point
of flaking.
That’s it
Vitamin D
Lecture 3: Minerals
Jan 2020 Vitamin D is a collective term used to refer to any one of
4 similar substances, only one of which has biological
activity:
• ergocalciferol (D2 from plants);
• cholecalciferol (D3 from animals);
Melford John PhD • 25-hydroxyvitamin D (circulating form);

Biochemistry Unit
Department Of Preclinical Sciences • 1,25-dihydroxyvitamin D (calcitriol, active form).
Vitamin D Vitamin D
Vitamin D different from other vitamins:
• synthesised in sufficient amounts in sunlight; • Facilitates intestinal absorption of Ca
• there are 4 forms, only one of which is active; • Necessary for growth of strong bones and teeth
through the proper utilisation of Ca and P
• functions like a hormone;
• Protects against cancer
• it follows a circuitous route in the body involving
the skin, the liver, and the kidney;
• significant amounts stored in fat cells.
Food sources of vitamin D Vitamin D

• Dietary vitamin D provided primarily by foods of animal
origin: synthesis
liver, beef, veal, eggs (mainly yolk), milk, cheese,
butter, herring, salmon, tuna, and sardines.
• Some foods fortified with vitamin D
• Vitamin D not prone to cooking or storage loss

Vitamin D
Lack of vitamin D
Required Dietary Allowance:
• A lack of vitamin D is very common
• children: 400 IU/day;
• adults: 800 IU/day; • Most food contain very little
• pregnancy and lactation: 1000 IU/day. • A mild lack of vitamin D causes tiredness and
general aches and pains
Vitamin D deficiency Vitamin E

• Potent antioxidant that prevents oxidation of fatty acids
• In children causes rickets, a disorder manifested by • Protects lipids in membranes from oxidative damage:
bowed legs particularly phospholipids of mitochondria and
endoplasmic reticulum, which are more unsaturated;
• In adults causes osteomalacia
cell membranes of lung, brain, and erythrocytes also
• In the elderly can also cause loss of hearing susceptible.
may be related to malformation of the middle ear

bones or changes that occur to the bones.
RDA of vitamin E Sources of vitamin E
• In it's various forms found in plants and animals

• Plant oils are a superior source:
• adult male : 30 IU /day;
• female : 25 IU/day; canola, olive, sunflower, corn, soybean, peanut
butter, whole-grain cereals, legumes.
• child : 10-20 IU/day.
• Fatty tissue of animals
Vitamin E
Vitamin E deficiency
Protects erythrocytes from hemolysis by oxidising agents
Deficiency in humans is quite rare
Required for normal reproduction in animals
Prevents liver necrosis and muscular dystrophy Muscular weakness
Fragile red blood cells
Vitamin K Vitamin K
A group of 3 structurally similar vitamins Sources:
Required for the maintenance of normal concentration of • green leafy vegetables and tomatoes;
blood clotting factors
• synthesised by microorganisms in the intestinal
Some studies suggest they help maintain strong bones in the tract.
elderly
Vitamin K Vitamin C
Deficiency: Involved in the hydroxylation of proline and lysine

residues:
• excessive bleeding.
• required for synthesis of collagen.
Helps in the absorption of iron
Acts as an antioxidant
Vitamin C Vitamin C
Sources:
• citrus fruits;
• adults: 60 mg/day;
• tomatoes;
• strawberries; • children: 40 mg/day.
• green vegetables;
• guava.
Vitamin C deficiency Minerals

Minerals fulfil the following functions:
• co-factors of enzymes;
• electrolytes;
• structure of tissues such as: bones, teeth,
nails, blood, nerves cells, and muscles.
Minerals of the body Dietary intake of minerals

• Most people eating a mixed diet are likely to
receive adequate intakes of minerals
• Amounts required vary:
g per day for Na and Ca;
mg per day for Fe and Zn;
µg per day for trace elements.
Calcium Calcium bank
Most abundant mineral
• Bones are a bank from which Ca
99% of the body’s Ca exists in bones and teeth: may be drawn or returned
• provide structure and strength; • In times of need:
• serve as a reserve bank that can release Ca when Ca absorption from the intestine
blood level drops. is increased;
1% of Ca in fluids that bathe and fill cells loss from the kidneys is reduced.
Calcium Osteoporosis
Role of Ca: • Osteoporosis, or adult bone loss, occurs if a
• regulation of ion transport; person’s Ca bank is not sufficient
• maintenance of blood pressure; • A diet low in Ca during the growing years may
• blood clotting; prevent the achievement of peak bone mass
• muscle contraction;
• secretion of hormones, digestive enzymes, and
neurotransmitters;
• activation of cellular enzymes.
Calcium loss with age Bones and calcium loss

• Bone loss is an inevitable consequence of ageing
• It can be reduced by a good diet and exercise
Bones and calcium loss Hypocalcemia
• Obtaining enough Ca in childhood helps Low blood Ca causes tetany, characterised by:
ensures the skeleton starts adulthood with a • paresthesias (tingling, burning, pricking, or numbness) of
the lips, tongue, fingers and feet;
high bone density
• carpopedal spasm, which may be prolonged and painful;
• Most girls in their bone-building years fail to • generalised muscle aching;
meet their Ca needs • spasms of facial musculature.
Although Ca is typically measured in labs from

blood plasma, tetany is caused by low ionic Ca in
extracellular and intracellular fluid
Phosphorous Magnesium
• 85% of body’s phosphorus found An activator of many enzymes especially those involving
combined with Ca in bones and teeth transfer of phosphate groups from ATP such as:
• hexokinase;
• Phosphorus also:
• phosphofructokinase.
helps maintain acid-base balance;
is part of nucleic acids (DNA, RNA); Mg also activates a number of enzymes such as:
involved in energy metabolism (ATP, GTP); • enolase;
is part of cell membranes (phospholipids). • glucose-6-P dehydrogenase;
• pyruvate carboxylase;
• thiokinase;
• glucose 6 phosphogluconate dehydrogenase.
Magnesium Sodium
• The major cation of the extracellular fluid
• Most of the body’s Mg is in the bones • Roles:
• Can be drawn out of cells for use • maintenance of electrolyte balance of body fluid;
• essential for nerve transmission;
• involved in active transport of glucose, galactose, and
amino acids across intestinal mucosa.
Sodium Potassium
30 to 40 % of body’s Na on surface of bone • Principal positively charged ion inside cells
crystals where it is easily drawn upon to
• Plays major role in maintaining electrolyte
replenish blood concentrations
balance
Chloride
Na, K and blood pressure
• Cl is the body’s major negative ion:
• Low K intake raises blood pressure
responsible for stomach acidity.
• High K intake appears to reduce hypertension
• No known diet lacks Cl
• High Na intake raises blood pressure
Trace minerals Iodine

• Hard to determine precise roles in man as it is
difficult to conduct experiments with diets • Iodine is part of thyroxine:
lacking a single element hormone responsible for regulating basal
metabolic rate.
• Studies are generally done in laboratory animals
that are fed controlled diets • Iodine in plants varies depending on the soil in
which they are grown
• Iodine in animals varies depending on the source
of the food they eat
Iodine deficiency Iron
• Goitre – cells of the thyroid gland enlarge until it Necessary for synthesis of Fe containing
makes a visible lump in the neck proteins:
• Cretinism – severe iodine deficiency during pregnancy • heme proteins:
causes death of foetus or cretinism: haemoglobin, myoglobin, catalase and cytochrome.
irreversible mental and physical retardation; • non-heme proteins:
ferritin, transferrin, aconitase and sucinate
a common and preventable cause of mental retardation.
dehydrogenase.
Iron-deficiency anaemia Causes of iron deficiency and anaemia

Symptoms:
• fatigue and weakness;
• Globally the most common nutrient deficiency
• pale skin and mucous ( >1.2 bn people)
membranes;
• Usually caused by:
• rapid heartbeat or a new heart
murmur; not eating sufficient food;
• irritability; consumption of the wrong type of foods.
• decreased appetite;
• dizziness or a feeling of being
lightheaded.
Zinc Problem: too little zinc

Zn required by nearly 100 enzymes e.g.:
carbonic anhydrase;
alkaline phosphatase;
liver alcohol dehydrogenase;
carboxyl peptidase A;
DNA polymerase;
Cytosolic superoxide dismutase.
Required for making testosterone and healthy sperm
Boy is 17 years old but is only 4 feet tall. Genitalia like those
of a six year old. Caused by Zn deficiency.
Selenium Glutathione
• Selenoperoxidases involved in antioxidant systems
• Se is an integral component of glutathione
peroxidase:
2 glutathione + H2O2 -> oxidised glutathione + 2 H2O
• Glutathione peroxidase 2 (GPX2) in intestinal
epithelial cells participates in detoxification of
dietary organic hydroperoxides:
also present in liver.
Fluoride Chromium
• Cr works with insulin to control blood glucose
• F stabilises bones and makes teeth resistant to concentrations
decay
• Cr is present in a variety of unrefined foods
• Excess F discolours teeth
• It is estimated that 90 % of US adults consume
• Large doses of F are toxic
less than the recommended minimum intake of 50
micrograms a day
Copper Copper
• Cu is needed to form haemoglobin and Cu containing enzymes:
collagen and assists in many other body • ceruloplasmin;
• cytochrome oxidase;
functions • dopamine oxidase;
• Deficiency is rare • monoamine oxidase and diamine oxidase;
• cytoplasmic superoxide dismutase;
• lysyl oxidase involved in the cross-linking process in the
conversion of tropocollagen to collagen;
• tyrosinase of the melanin synthetic pathway.
Manganese Other trace minerals
Acts as a cofactor for enzymes such as: Many other trace elements play important roles
• acetyl CoA carboxylase; in the body e.g.:
• mitochondrial superoxide dismutase;
• arginase; molybdenum, boron, cobalt, nickel, silicon.
• 6 Phosphate-gluconate dehydrogenase;
• squalene synthetase;
• isocitrate dehydrogenase;
• glutamine synthetase;
• kinases;
Summary Summary
Mineral deficiencies Mineral toxicity

• Occur when foods come from one region where Generally too much of any mineral can be toxic:
the soil is deficient in one or more minerals • too much Se is toxic;
• Deficiencies of iodine and selenium, occur in • Zn in high doses can cause serious illness or
many areas of the world even death;
• Fe supplements are a leading cause of fatal
accidental poisonings among US children
under six years old.
That’s it?
61
Lecture 4: Assessing Nutritional Status & Nutrition-Related Diseases Assessment of nutritional status
Jan 2020
Benefits:
• immediate treatment;
• development of healthcare programmes to meet community needs;
• assessment of effectiveness of programmes;
Methods of nutritional assessment:

• direct methods;
• indirect methods.
Melford John PhD

Biochemistry Unit
Department Of Preclinical Sciences
1 2
Causes of deaths
Deaths from NCDs
What kills more people, infectious diseases or
noncommunicable diseases? What % of deaths are caused by NCDs?
• high-income countries 87%;
• upper-middle income countries 81%;
• lower-middle income countries 56%;
• lower low-income countries 36%.
The richest countries in the world 13/01/2017, 18:41
12 Switzerland 59,376 Europe 83.58 02 (7.509)
United North
13 57,294 79.57 13 (7.104)
Source: World Health Statistics by WHO
States 2013 America
14 Saudi Arabia 54,078 Asia 74.79 34 (6.379)

3 4 15 Netherlands 50,846 Europe 82.05 07 (7.339)
16 Bahrain 50,303 Asia 77.05 42 (6.218)
17 Sweden 49,678 Europe 82.76 10 (7.291)
The richest countries in the world The richest countries in the world 13/01/2017, 18:41
Australia 13/01/2017, 18:41
18 Australia 48,806 and 83.10 09 (7.313)
Oceania
MOST TRADED CURRENCIES survey). MOST TRADED CURRENCIES survey).
19 Germany 48,190 Europe 81.54 16 (6.994)
GLOBAL EDUCATION Lust for travel with family GLOBAL EDUCATION Lust for travel
20 Iceland 48,070with family83.13
Europe 03 (7.501)
WORLD'S TOP UNIVERSITIES WORLD'S TOP UNIVERSITIES
21 Austria 47,856 Europe 82.06 12 (7.119)
BEST US ONLINE COLLEGES BEST US ONLINE COLLEGES
22 Taiwan
List of the 50 richest47,790 Asia
countries ranked 79.84
by GDP 35 (6.379)
based on
FREE ONLINE COURSES List of the 50 richest countries ranked by GDP based onFREE ONLINE COURSES
purchasing-power-parity (PPP) per capita purchasing-power-parity
23 Denmark 46,603(PPP) per capita
Europe 80.74 01 (7.526)
Trinidad & Tobago

GDP(PPP) Average Happiness GDP(PPP) North Average Happiness
per capita life Ranking 24
Rank
Canada
Country
46,240
per capita America
Continent
82.63
life 06 (7.404)
Ranking
Rank Country Continent 2016 expectancy Rank 2016
2016 expectancy Rank 2016
25 Belgium (Int.$)
44,881 Europe (years)
81.43 (score)
18 (6.929)
(Int.$) (years) (score)
1 Qatar 129,727 Asia 78.68 36 (6.375)

1
26 Qatar
Oman 129,727
43,737 Asia
Asia 78.68
77.50 36no(6.375)
data
2 Luxembourg
United 101,936 Europe 82.30 20 (6.871)
2 Luxembourg 101,936 Europe 82.30 20 (6.871) 27 42,514 Europe 81.25 23 (6.725)
Kingdom
Are we a high income country?

3 Liechtenstein 89,400 Europe 82.38 no data
3 Liechtenstein 89,400 Europe 82.38 no data
The richest countries in the world
28 France 42,384 Europe 82.84 32 (6.478) 13/01/2017, 18:41
4 Singapore 87,082 Asia 83.69 22 (6.739)
4 Singapore 87,082 Asia 83.69 22 (6.739)
29 Finland
5 Brunei
https://www.countries-ofthe-world.com/richest-countries.html 41,813
79,711 Europe
Asia 81.47
79.49 05
no (7.413)
data Page 3 of 5
5 Brunei 79,711 Asia 79.49 no data

30 Japan
6 Monaco 38,894
78,700 Asia
Europe 84.08
89.67 53
no (5.921)
data
6 Monaco 78,700 Europe 89.67 no data
Equatorial
7 Kuwait 71,264 Asia 74.83 41no(6.239)
31 38,699 Africa 58.57 data
7 Kuwait 71,264 Asia 74.83 41 (6.239) Guinea
8 Ireland 69,375 Europe 81.52 19 (6.907)
8 Ireland 69,375 Europe 81.52 19 (6.907) 32 South Korea 37,948 Asia 82.75 58 (5.835)
9 Norway 69,296 Europe 82.11 04 (7.498)
9 Norway 69,296 Europe 82.11 04 (7.498) 33 Malta 37,891 Europe 81.16 30 (6.488)
United Arab
United Arab 10
34 Emirates
Andorra 67,696
37,200 Asia
Europe 77.55
82.72 28no(6.573)
data
10 67,696 Asia 77.55 28 (6.573)
Emirates Australia
11 San Marino 64,443 Europe 83.53 no data
The richest countries in the world
11 San Marino 64,443 Europe 83.53 no data 35 New Zealand
13/01/2017, 18:41
37,108 and 82.42 08 (7.334)
Oceania
https://www.countries-ofthe-world.com/richest-countries.html Page 2 of 5
12
https://www.countries-ofthe-world.com/richest-countries.html
Switzerland 59,376 Europe 83.58 02 (7.509)
36Page 2Spain
of 5 36,451 Europe 83.22 37 (6.361)
United North
13 57,294 79.57 13 (7.104) 37 Italy 36,313 Europe 83.75 50 (5.977)
States America
38 Israel 34,834 Asia 83.01 11 (7.267)
14 Saudi Arabia 54,078 Asia 74.79 34 (6.379)
39 Cyprus 34,387 Europe 80.75 69 (5.546)
15 Netherlands 50,846 Europe 82.05 07 (7.339)
Czech
16 Bahrain 50,303 Asia 77.05 42 (6.218) 40 33,223 Europe 79.10 27 (6.596)
Republic
17 Sweden 49,678 Europe 82.76 10 (7.291) 41 Slovenia 32,028 Europe 80.98 63 (5.768)
Australia Trinidad and North

18 Australia 48,806 and 83.10 09 (7.313) 42 31,934 70.74 43 (6.168)
Tobago America
Oceania
43 Slovakia 31,182 Europe 76.67 45 (6.078)
19 Germany 48,190 Europe 81.54 16 (6.994)
The richest countries in the world 13/01/2017, 18:41
44 Lithuania 29,882 Europe 73.72 60 (5.813)
20 Iceland 48,070 Europe 83.13 03 (7.501)
45 Estonia 29,502 Europe 77.20 72 (5.517)
21 Austria 47,856 Europe 82.06 12 (7.119)
46
https://www.countries-ofthe-world.com/richest-countries.html Portugal 28,515 Europe 81.72 94 (5.123) Page 4 of 5
22 Taiwan 47,790 Asia 79.84 35 (6.379)
47 Seychelles 28,148 Africa 73.75 no data
23 Denmark 46,603 Europe 80.74 01 (7.526)
48 Poland 27,715 Europe 77.98 57 (5.835)
North
24 Canada 46,240
America
82.63 06 (7.404)
49 Malaysia 27,234 Asia 75.33 47 (6.005)
25 Belgium 44,881 Europe 81.43 18 (6.929)

50 Hungary 27,211 Europe 75.58 91 (5.145)
5 26 Oman 43,737 Asia 77.50 no data
6
United
27 Kingdom
42,514 Europe 81.25 23 (6.725)
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28 France 42,384 Europe 82.84 32 (6.478) © 2008–2017, www.countries-ofthe-world.com
https://www.countries-ofthe-world.com/richest-countries.html Page 3 of 5
Obesity update - © OECD 2017
se Figure 1: Obesity among adults, 2015 or nearest year
3.7 Japan
Self-reported data 5.3 Korea
Women Obesity update - © OECD 2017
Measured data 9.8 Italy Men
10.3 Switzerland Rising overweight rates in adults 15 to 74 years
12.0 Norway Figure 2: Rising overweight (including obesity) rates in adults aged 15-74 years
12.3 Sweden
12.8 Netherlands
14.7 Austria 80%
14.9 Denmark
Obese
Rates of overweight (including obesity)

15.3 France
Mexico
16.3 Slovak Rep.
70%
countries 16.6
16.7
Portugal
Poland
United States
16.7 Spain
17 Greece
60% Hungary
17.8 Israel
18 Estonia
England
18.6 Belgium
Canada
19 Iceland 50%
19.2 Slovenia Spain
19.5 OECD
21 Czech Rep. France
21.3 Latvia 40% Italy
22.3 Turkey
22.6 Luxembourg Switzerland
23 Ireland
30%
n 23.6
24.8
Germany
Finland
Korea
25.1 Chile
25.8 Canada
26.9 United Kingdom 20%
27.9 Australia 1972 1976 1980 1984 1988 1992 1996 2000 2004 2008 2012 2016
30 Hungary
30.7 New Zealand
32.4 Mexico Year
38.2 7 States
United 8
Note: Overweight and obesity rates designate overweight and obesity prevalence rates. Age- and gender-adjusted rates of
5.0 India overweight (including obesity), using the 2005 OECD standard population. Measured height and weight in England, Hungary,
5.7 Indonesia Korea, Mexico and the United States; self-reported in other countries.
7.0 China Source: OECD analysis of health survey data.
17.3 Lithuania
19.6 Russian Fed.
20.8 Brazil
20.9 Colombia
24.4 Costa Rica Figure 3: Self-reported overweight (including obesity) in children aged 15 years
26.5 South Africa
40 30 20 10 0 0 10 20 30 40 2001-02 2013-14
Indirect methods
% of population aged 15 years and over % of population aged 15 years and over 35
Direct methods 30
31
Source: OECD (2017), OECD Health Statistics 2017 (Forthcoming in June 2017).
www.oecd.org/health/health-data.htm
Note: The statistical data for Israel are supplied by and under the responsibility of the relevant Israeli authorities. The use of such data
by the OECD is without prejudice to the status of the Golan Heights, East Jerusalem and Israeli settlements in the West Bank under the
terms of international law. 25
Use community health indices that reflects nutritional
24.5
Deal with the individual and measure objective criteria 20
influences
21.5
3 %
18
17
17
17
15
16.5
16.5
Summarised as ABCD:
16
16
16
15.5
15.5
15.5
15.5
15
15
These include:
14
14
14
13
A. anthropometric methods;
12.5
12.5
12.5
12.5
12.5
12
12
10
12
• ecological variables including crop production;
10.5
9.5
B. biochemical methods; 5
• economic factors e.g. per capita income, population;
C. clinical methods;
D. dietary evaluation methods. 0 density and social habits;
• vital health statistics particularly infant under 5
* mortality and fertility index.
*
Note: * Data for 2009-10. Child overweight is defined with IOTF age- and gender- specific BMI cut-offs.
Source: Currie, C. et al. (2004); Inchley et al. (2016).
9 10
A. Anthropometric measurements A. Anthropometric measurements

• The measurement of body weight and proportions 1.Height/age ratio and weight/height
2.Mid-upper arm circumference

• An essential part of clinical examination of infants,
children, and pregnant women 3. Head circumference
• Evaluates both malnutrition and nutritional excesses 4.Skin fold thickness
5.BMI
• Assesses current nutritional state
6.Waist/hip ratio
11 12
A1. Height/age ratio A2. Mid upper arm circumference
• Accurate measurement of height and weight is necessary to • Measured half-way between the acromion process of
evaluate the proper growth of a child the scapula and the tip of the elbow (ulnar) with the
arm hanging vertically and forearm supinated
• Ht/Age:
appropriate height for given age. • Provides estimate of arm muscle area:
reflects skeletal protein reserves.
• Wt/Ht:
appropriate weight for given height.
13 14
A3. Head circumference A4. Skin fold

Triceps, biceps, sub-scapular, supra-iliac used in
Useful in children under the age of 3 and is an indicator of combination to estimate body fat
non-nutritional abnormalities
Malnutrition will have to be severe to affect head

circumference
Estimates body fat using software
15 16
A5. Body mass index (BMI)

A5. Body mass index (BMI)
High BMI (obesity level) associated with:
• type 2 diabetes;
• high risk of cardiovascular morbidity and mortality.
BMI = Weight (kg) / Height 2 (m)
17 18
A6. Waist/hip ratio A6. Hip circumference
• Measured at the point of greatest circumference around
Interpretation of waist/hip hips and buttocks to the nearest 0.5 cm
Ratio (WHR)
• The subject should be standing and the measurer
High risk WHR: should be squat beside him/her
> 80% for females; • Both measurement should be taken with a flexible, non-
> 95% for males. stretchable tape in close contact with the skin, but
without indenting the soft tissue.
Indicates central obesity
High risk of diabetes and Hip To Waist Ratio
cardiovascular disorders. An Important Parameter
19 20
A6. Waist circumference Advantages of anthropometry

• Simple and non-invasive
Waist circumference predicts mortality better than any other • Equipment is inexpensive and portable
anthropometric measurement
• Relatively unskilled personnel can perform measurements
Male Female
• Methods are reproducible
Level 1 >94 cm >80 cm
• Measures long term nutritional history
Level 2 >102 cm >88 cm
• Quickly identifies mild to moderate malnutrition
• level 1 is the maximum acceptable waist circumference • Measures many variable of nutritional significance like
• level 2 highlights obesity and requires weight management height weight, skin fold thickness, head circumference
waist and hip ratio and BMI
21 22
Disadvantages of anthropometry B. Biochemical methods

• Relatively insensitive to short term nutritional status
• Cannot identify specific nutrient deficiencies • Haemoglobin gives a useful indication of the overall state
• Some measurements such as skin-folds difficult to carry of nutrition
out for obese people • Beside anaemia it also gives idea about protein and trace
• Ethnic differences in fat deposition element nutrition
• HbA1c Glycation test gives average blood glucose levels
over the past 3 to 4 months
• Urine examination for albumin and sugar
• Levels of lipoproteins and triglycerides in the blood
23 24
B. Biochemical methods B. Glycation of proteins
Advantages of Biochemical Measurements: • Elevated glucose levels associated with increased
• able to detect early changes in body metabolism and nutrition glycation and cross-linking of proteins:
before the appearance of clinical signs;
increases risk of type 2 diabetes and coronary heart
• accurate and reproducible; disease.
• useful to validate data obtained from dietary methods (e.g.
comparing salt intake with 24- hour urinary excretion. • Fructose x7 to x10 higher glycating agent than glucose
Disadvantages of Biochemical Measurements:

• expensive;
• time consuming;
• needs trained personnel and facilities.
25 26
B. Glycation of proteins B. HbA1c glycation test

Megerssa and Tesfaye 19
Figure 1. Chemistry of protein glycation adopted from characterization of glycation sites on human serum albumin using mass
27 28
spectrometry: PhD Dissertation by Omar St. Aubyn Barnaby Graduate College at the University of Nebraska, 2010.
forward reaction rate. The second main determinant is (Brutis et al., 2006). Glycation occurs at several amino
the duration of a p o ein exposure to an elevated glucose acid residues of the different variants of hemoglobin
concentration, which relates to both its residence time in (HgbA, HgbA2 and HgbF) and results in a product called
the circulation and episodes of hyperglycemia (Schleicher glycated hemoglobin (GHb). Chromatographic analysis of
and Wieland, 1986). Hyperglycemia is associated with a HgbA reveals a number of minor species HgbA1a, HgbA1b
defect in insulin secretion, insulin action, or both. Insulin and HgbA1c that are collectively known as fast hemoglobin.
C. Clinical methods
deficiency in turn leads to diabetes mellitus with
disturbances of carbohydrate, fat and protein metabolism
HgbA1c is the specific amadori product of glucose
conjugated with valine at the N-terminal of both chainsFull of vitamins?
(Megerssa et al., 2013). of HgbA. This product accounts for approximately 80% of
In vivo, the amount of any glycated protein maintained HgbA1 and about half of the total GHb (Benjamin and
in a steady state and is influenced by time-averaged Sacks, 1994). Other glycohemoglobin species products
Examination
glycemia, the of: rate of glycation, and the rate of the have glucose linked to an -amino group of one or more
p• o hair,
ein nails, skin,
deg ada iongums, eyes,almuscles,
o emo f om hetongue,
ci c lamouth,
ion. bones
of theirand
lysine residues on their or chain (Cohen and
During the process of glycation (Figure 1), glucose Clements, 1999).
joints, and thyroid gland.
adducts can continue to form until equilibrium is reached, Formation of GHb is essentially irreversible and its
and a new population of the protein is subjected to the concentration in blood depends on both the life span of
pertinent reaction kinetics as an older population is
Advantages: red cells and blood glucose concentration. As the rate of
physiologically removed (Schleicher and Wieland, 1986). formation of GHb is directly proportional to the concen-
• fast
In and easy
persistent to perform;;
hyperglycemic patients, hemoglobin (Hgb), tration of glucose in the blood, the GHb concentration
• inexpensive;
albumin, lipoproteins and other tissue proteins are non- represents the integrated values for glucose in the
enzymatically glycated to a great extent and are hall preceding 6 to 8 weeks. This provides an additional
• non-invasive.
marks of diabetes mellitus. Hence, glycated hemoglobin criterion for assessing glucose control because GHb
(GHb) and glycated albumin (fructosamide) are widely values are free of day-to-day glucose fluctuations and are
accepted as reliable indicators of metabolic control in
Limitations: unaffected by recent exercise and food ingestion (Brutis
diabetic patients (Kobayashl et al., 1990). et al., 2006).
• may not detect early stages malnutrition.
CLINICALLY RELEVANT GLYCATED PROTEINS Clinical use of HbA1c
Glycated hemoglobin 29 HbA1c level reflects the blood glucose level consistent 30
with the live span of red blood cell, which is close to 120
In adults, Hgb is usually constituted of HgbA (97%), days (Nitin, 2010). Hence, higher levels of HbA1c are
HgbA2 (2.5%) and Hgb F (0.5%). HgbA, in turn, is made found in people with persistently elevated blood sugar, as
C. Signs of nutritional deficiency - hair C. Signs of nutritional deficiency - mouth
Mouth Symptons Deficiency
Hair Symptons Deficiency

Glossitis Riboflavin (B2) and niacin (B3)
Angular stomatitis cheilosis Riboflavin, pyridoxine (B6), and niacin

Hair loss Zn deficiency and fissured tongue
Easy to pull out Protein deficiency Bleeding and spongy gums Vitamin C and/or A
Corkscrew coiled hair Vitamin C and/or A deficiency
Leukoplakia Vitamin A, cobalamin (B12), folic acid
(B9), and niacin
General deficiency Pyridoxine, niacin, and iron
31 32
C. Signs of nutritional deficiency - eyes C. Signs of nutritional deficiency - nails

Nail Symptons Deficiency
Eye Symptons Deficiency
Spooning Iron deficiency

Night blindness Vitamin A
Transverse lines Protein deficiency

Photophobia, blurring and
conjuctival inflammation Vitamin A
spooning
33 34
C. Signs of Nutritional deficiency - skin C. Pallor

Skin Symptons Deficiency
Pallor Folic acid, iron, and cobalamin (B12)
Follicular hyperkeratosis Vitamin A, C and E
Flaking dermatitis Cobalamin, A, Zn, and niacin (B3)
Pigmentation and Niacin & protein energy malnutrition

desquamation
folic acid (B9)
Bruising purpura
cobalamin (B12)
Vitamin K, C, and folic acid (B9)
iron
35 36
C. Desquamation C. Purpura
vitamin K
vitamin C
folic acid
niacin and
protein energy
malnutrition
37 38
C. Signs of nutritional deficiency - thyroid gland
Thyroid Gland Sympton Deficiency
Goitre Iodine deficiency Severe dietary deficiencies
39 40
Severe dietary deficiencies Severe acute malnutrition

• If dietary deficiencies are persistent, children stop growing: • Case fatality rate up to 21% without effective
intervention
low height for age;
• Includes two main clinical forms:
chronic malnutrition.
severe wasting (marasmus);
• If child experience weight loss or wasting:
nutritional edema (kwashiorkor).
low weight for height;
acute malnutrition.
• Clinical analysis determines if treatment will be in
hospital or with therapeutic ready-to-use-foods (RUFs)
• Both chronic and acute malnutrition may be further
at home
classified as moderate or severe
• Most children given therapeutic RUF treatment at home
41 42
Protein-calorie malnutrition Nutritional deficiency
• Protein-calorie malnutrition reflects starvation or specific
deficiencies
• A deficiency of proteins in diets relatively high in

carbohydrates leads to kwashiorkor
• A deficiency of all components of a diet leads to marasmus
• 4 to 5 million children younger than 5 die every year from

malnutrition
kwashiorkor marasmus
Carbs but no protein General deficiency

43 44
Kwashiorkor Protein-energy malnutrition

Derived from a term that means "the disease that occurs Marasmus characterised by:
when the next baby is born" • severe weight loss and wasting;
• subnormal body temperature;
Kwashiorkor is characterised by:
• decreased pulse and metabolic rate;
• edema;
• loss of skin turgor;
• low capillary-filtration rate;
• constipation;
• hypoalbuminemia;
• dermatitis.
• starvation diarrhea, consisting of frequent, small,
mucus-containing stools.
General deficiency
Carbs but no protein

45 46
Marasmus & kwashiokor Learning objectives

General deficiency Carbs but no protein
• Describe the components of the diet in terms of
macro and micronutrients
• Explain the term essential nutrients, and outline the

biochemical roles of essential nutrients
• Discuss the concept of recommended daily allowances

(RDA) for energy and essential nutrients, the
derivation of RDA values, and their limitations
• Describe the factors that determine energy and

protein requirements in man and animals
• Describe and compare the major contributors to

dietary energy intake between man and animals.
47 48
Summary
49
NITROGEN METABOLISM
LECTURE 2
WELCOME BACK
1. A young lady weighing 50 kg had a daily protein intake of 42 grams. Her intake was;
a. well above the daily requirement

b. just above the daily requirement
c. Just below the daily requirement
d. Well below the daily requirement
2. A patient had a nitrogen intake of 95 mg. He retained 40 mg while 15 mg was undigested. The
Biological value was;
a. 105 %
b. 80 %
c. 50 %
d. 25 %
Recall
Therefore intake required = 50 x 0.8

= 40 g
Answer b
Some cancers interfere with this process, interrupting the ‘programmed cell
death’ while allowing uncontrolled propagation of cancer cells
Aspartic acid Oxaloacetate
Glutamic Acid α-Ketoglutarate
NITROGEN
METABOLISM
LECTURE 3
Q. The diagram below reflects the early fate of an amino acid (W) in the presence of its
transaminase. It can be concluded that:
a. enzyme 1 is glutaminase
b. amino acid W is alanine
c. X must be pyruvate
d. enzyme 2 is inhibited by ATP
Q. The diagram below reflects the early fate of an amino acid (W) in the presence of its transaminase. It can
be concluded that:
a. enzyme 1 is glutaminase
b. amino acid W is alanine
c. X must be pyruvate
d. enzyme 2 is inhibited by ATP
Can’t be (a); glutaminase is not a transaminase as stated in the question. Also free NH4 would
Be produced.
Can’t be (b) as pyruvate would be a product
Can’t be (c) as one of the two products must be Glu given that a–KG was a starting material
Answer is (d). Glutamate dehydrogenase is inhibited by ATP (check notes)

IMPORTANT CHEMISTRY
KIDNEY: Uses NH3 To Control the pH of Urine
G.S is glutamine synthetase\synthase

MUSCLE-THE ALANINE CYCLE
AMINO ACID BIOSYNTHESIS
AMINO ACID RELATED DISEASES
1. AMINO ACID URIAS
Proximal tubule of
Kidneys fails to
reabsorb amino
acids.
In cystinuria Cystine
and three dibasic A.A
( Ornithine, Arginine
and Lysine) COAL are
not reabsorb.
Elevated in urine.
Causes kidney stones

HISTIDINE
Recall the role of NADH in ATP
synthesis
Thus no NADH; no Energy leading

to disease.
PELLAGRA (The 3D’s)

Dermatitis
Dementia
Diarrhea
TRYPTOPHAN (In Carcinoid Syndrome)
THANK YOU
VERY MUCH
Objectives
At the end of this this topic you should be able to:
• List the risk factors associated with cardiovascular

diseases
• Explain how some of the risk factors of CVD can be
modified with proper life style and diet
Risk Factors for Cardiovascular
Disease
Cardiovascular diseases are the leading cause of illness
and death in the world
The majority of cases stem from atherosclerosis
In coronary heart disease:

The arteries to the heart muscle (myocardium) are
narrowed leads to reduced blood supply to the heart can
result in chest pain (angina pectoris)
If a narrowed blood vessel is completely blocked by a
blood clot, the area of the heart just beyond the blockage
is denied oxygen and nourishment, resulting in a heart
attack (MI) Dr.S Nayak 2
Ten risk factors help to predict the likelihood of CHD are
1. Heredity
2. Gender
3. Age
4. Cigarette smoking
5. High blood pressure
6. Diabetes
7. Obesity
8. Lack of physical activity
9. Abnormal blood cholesterol
10.Homocysteine levels
The more risk factors, the greater the likelihood of developing
heart disease.
Heredity, gender, and age cannot be modified, but the others
can be influenced by the individual's
Dr.S Nayak behavior 3
Major risk factors [can't be changed]
• Age — Over 83 percent of people who die of coronary heart
disease are 65 or older
• Gender — Men have a greater risk of heart attack than

women do.
• Heredity (including Race) — Children of parents with heart

disease are more likely to develop it themselves.
African Americans have more severe high blood pressure
than Caucasians and a higher risk of heart disease.
This is partly due to higher rates of obesity and diabetes.

Most people with a strong family history of heart disease
have one or more other risk factors.
Dr.S Nayak 4
Major risk factors [can be modified]
Can be modified through treatment or control by
changing lifestyle or taking medicine
• Tobacco smoke
Smokers' risk of developing coronary heart disease
is 2–4 times that of nonsmokers. Cigarette smoking
is a powerful independent risk factor for sudden
cardiac death in patients with coronary heart
disease.
• High blood cholesterol

Risk of coronary heart disease increases when blood
cholesterol rises.
When other risk factors (high BP and tobacco
smoke) are present, this risk increases even more.
Dr.S Nayak 5
Total blood cholesterol is classified by levels:
Desirable: under 200 mg/dL
Borderline: 200-239 mg/dL
High risk: 240 mg/dL and above
• LDL Cholesterol
Optimal: Less than 100 mg/dL
Near or above optimal: 100-129 mg/dL
Borderline high: 130-159 mg/dL
High: 160-189 mg/dL (high risk)
Very high: 190 mg/dL and above (very high risk)
• HDL Cholesterol
Major heart disease risk factor: less than 40 mg/dL
Protection against heart disease: 60 mg/dL and Above
Dr.S Nayak 6
• High blood pressure (BP)
High BP increases the heart's workload, causing the
heart to thicken and become stiffer. It also increases
stroke, heart attack, kidney failure and congestive heart
failure. When high BP exists with obesity, smoking, high
blood cholesterol levels or diabetes, the risk of heart
attack or stroke increases several times.
Healthy adult [at rest] should have a systolic pressure
below 120 and a diastolic pressure below 80
Dr.S Nayak 7
• Physical inactivity
Inactive lifestyle is a risk factor for coronary heart
disease. Regular, moderate-to-vigorous physical
activity helps prevent heart and blood vessel
disease. Physical activity can help control blood
cholesterol, diabetes and obesity
Dr.S Nayak 8
• Obesity and overweight
People with excess body fat — especially at the waist
are more likely to develop heart disease and stroke
even if they have no other risk factors.
Excess weight increases blood pressure and blood
cholesterol and triglyceride levels, and lowers HDL
cholesterol levels. It makes diabetes more likely to
develop
Dr.S Nayak 9
 High Homocysteine
The blood level of homocysteine is 15 micromoles/L.
Increased level associated with cardiovascular
disease
• Diabetes mellitus
o Increases the risk of developing cardiovascular
disease.
o Increases the risk of heart disease and stroke.
Uncontrolled blood sugar increases risk more.
Dr.S Nayak 10
About three-quarters of people with diabetes die of
some form of heart or blood vessel disease. It is
important to work with healthcare provider to manage it
and control any other risk factors you can.
Other factors contribute to heart disease risk

• Stress may be a contributing factor. For example,
people under stress may overeat, start smoking or
smoke more than they otherwise would.
• Too much alcohol can raise blood pressure, cause

heart failure and lead to stroke.
It can contribute to high TG and obesity.
Dr.S Nayak 11
Experts say that moderate intake is an average of one to
two drinks per day for men and one drink per day for
women.
One drink is defined as 1½ fluid ounces (fl oz) of 80-proof

spirits (such as Scotch, vodka, gin, etc.), 1 fl oz of 100-
proof spirits, 4 fl oz of wine, or 12 fl oz of beer.
But drinking more than a moderate amount of alcohol can

cause heart-related problems such as high blood
pressure, stroke, irregular heartbeats, and
cardiomyopathy (disease of the heart muscle)
Ref: Essentials of Biochemistry

Dr.S Nayak 12
Tricarboxylic acid cycle (TCA Cycle)
[Kreb’s cycle] [Citric acid cycle]
Objectives
At the end of this topic the students should be able to:
1. Describe further processing of acetyl in TCA cycle

2. Calculate the energy yield per turn of the cycle
Prof S Nayak
Tricarboxylic acid cycle (TCA Cycle)
[Kreb’s cycle] [Citric acid cycle]
• Is the final common oxidative pathway for carbohydrates,

fats and amino acids
• Along with energy, cycle supplies many intermediates
required for the synthesis of amino acids, glucose, heme
etc.
• Site: mitochondrial matrix
• Oxidation of acetyl CoA  Co2 + H2O
• Occurs in a cyclic manner, generate ATP
• Two carbon, acetyl CoA + 4 carbon, Oxaloacetate
= 6 carbon tricarboxylic acid, citrate
Prof S Nayak
Prof S Nayak
Cis-aconitate is a transient one with very short half-life.
Immediate H2O added to it and forms Isocitrate\
CO2
 Isocitrate  oxalosuccinate -ketoglutatrate.
It is an oxidative decarboxylation
Oxalosuccinate is unstable so it undergoes spontaneous
decarboxylation to from  -KG
TCA is both catabolic and anabolic  amphibolic
Prof S Nayak
Energetics of TCA Cycle
Steps 4, 6, 10  3 NADH
1 NADH = 3 ATP] 3 ATP x 3 = 9 ATP
Step 8  1 FADH2
1 FADH2 = 2 ATP] 2 ATP x 1 = 2 ATP
Step 7  1 GTP
1 GTP = 1 ATP 1 ATP x 1 = 1 ATP
Therefore 1 acetyl CoA gives 12 ATP
Therefore 1 acetyl CoA gives 12 ATP
Two acetyl CoA in citric acid cycle produces 24 ATP
Energetics of complete oxidation of glucose
Aerobic glycolysis  8 ATP
Oxidation of 2 pyruvate = 6 ATP
Oxidation of 2 Acetyl CoA by TCA cycle  24 ATP
Net Gain = 38 ATP
Prof S Nayak
Amphibolic nature of TCA cycle
Non essential aa Aspartate Acetyl CoA
Purines , Transamination
Pyrimidines.
Oxaloacetate Citrate Acetyl CoA
Fatty acids,
steroids
Pyruvate Malate  -KG
Transanimation
Succinyl COA Glutamate
Heme Non-essential a a,
purines
Prof S Nayak
Anaplerosis
The reactions concerned to replenish the intermediates of

TCA cycle are called anaplerotic reactions or anaplerosis
Pyruvate +CO2 + ATP carboxylase oxaloacetate + ADP + Pi
Pyruvate +CO2 + NADPH + H + Malic enzyme Malate
Inhibitors that inhibit the enzymes of TCA cycle are:

Aconitase fluoroacetate
- Ketoglutarate DH Arsenite Non-competitive
Succinate DH Malonate }- competitive

Prof S Nayak
Regulation of TCA cycle
1. Citrate synthase: inhibited by ATP, NADH, acyl CoA

and succinyl CoA
2. Isocitrate dehydrogenase: Inhibited by ATP and NADH
and activated by ADP
3. -KG dehydrogenase inhibited by NADH & succinyl CoA
The availability of ADP: Important for proceeding the TCA

cycle
if not oxidation of NADH and FADH2 through election transport
chain stops. Accumulation of NADH and FADH2, inhibit the
enzymes of TCA cycle.

Prof S Nayak
BODY FLUIDS
Dr. Neetu Mohan
University of the West Indies, St. Augustine
OUTLINE
• Body fluid compartments
• Measurement of fluid in compartments
• Osmosis, osmolarity, osmotic pressure
• Measurement of solute concentrations
• Freezing point depression
• pH
DAILY INTAKE OF WATER
• Water is added to the body by:
1. Ingestion in the form of liquids or water in food

• Adds about 2100 ml/day to the body fluids
2. Synthesized in the body

• oxidation of carbohydrates (adds about 200 ml/day)
DAILY INTAKE OF WATER
Intake of water is highly variable among different people and even within
the same person on different days, depending on:
• climate
• habits
• level of physical activity
DAILY LOSS OF BODY WATER
(1) Insensible Water Loss.
continuous loss of water by evaporation (700
ml/day)
• from the respiratory tract (about 300 to 400
ml/day)
• Air must attain vapor pressure of about 47
mmHg before it is expelled
• vapor pressure of inspired air usually less
than 47 mm Hg ˂47mmHg 47mmHg
Water from lungs absorbed by air to
increase vapour pressure
• In cold weather, atmospheric vapor
pressure is nearly 0
greater loss of water from lungs (dry
feeling in the respiratory passages in
cold weather)
• diffusion through the skin (about 300

to 400 ml/day)
• minimized by cholesterol-filled
cornified layer of skin
• If cornified layer is denuded, (e.g
extensive burns), the rate of
diffusion can increase up to 10-
fold, to 3 to 5 L/day.
• burn victims given large
amounts of fluid, usually
intravenously, to balance fluid
loss.
• (2) Water Loss in Sweat
• highly variable
• physical activity
• environmental temperature
• normally about 100 ml/day
• in very hot weather or during heavy exercise, water loss in sweat can increase
to 1 to 2 L/hour)
• thirst mechanism activated to restore fluid balance
• (3) Water Loss in Feces
• small amount of water (100 ml/day) normally lost in feces
• Increases to several liters a day in people with severe diarrhea
• (4) Water Loss by the Kidneys
• urine excreted by kidneys
• can be as low as 0.5 L/day in dehydrated person or up to 20 L/day in a person
drinking large amounts of water
TOTAL BODY WATER (TBW)
• Total body water about 60% of the body weight,
or (42 liters for average 70kg adult human)
• Percentages change, depending on
• age – the older the person, the lower the
body fluid percentage
• aging associated with increased
percentage of the body weight being
fat ; fat decreases the percentage of
water in the body
• gender - women normally have more body
fat than men ; they contain slightly less
water than men of similar body weight.
• degree of obesity – people with high
percentage of body fat have less water
BODY FLUID COMPARTMENTS
• extracellular fluid
• interstitial fluid
• blood plasma
• transcellular fluid (small
quantity) (1-2L)
• fluid in the synovial,
peritoneal, pericardial,
intraocular spaces,
cerebrospinal fluid
• intracellular fluid
INTRACELLULAR FLUID COMPARTMENT (ICF)
• Fluid inside the 75 trillion

cells
• About 28 of the 42 liters of
fluid (67% of total body
fluid)
EXTRACELLULAR FLUID COMPARTMENT (ECF)
• All the fluids outside the cells
• about 14 liters (33% of total body
fluid) in an average 70-kilogram
adult.
• Interstitial fluid, > ¾ of the
extracellular fluid (about 11
liters)
• Plasma (noncellular part of
blood) ~ ¼ of the extracellular
fluid (about 3 liters)
• Transcellular fluid ~ 1-2 L
EXTRACELLULAR FLUID COMPARTMENT (ECF)
• Plasma exchanges substances

continuously with the interstitial fluid
• through pores of capillary
membranes
• these pores highly permeable to
almost all solutes in ECF (except
proteins)
• plasma and interstitial fluids have about
the same ionic composition except for
proteins, which have a higher
concentration in the plasma.
IMPORTANT CONSTITUENTS OF ECF
• ECF contains
• large amounts of Na+ and
Cl- ions
• reasonably large
amounts of HCO3- ions
• small quantities of K+ ,
Ca2+ , Mg2+ , PO43-, and
organic acid ions
IMPORTANT CONSTITUENTS OF THE
INTRACELLULAR FLUID
• ICF separated from ECF by cell
membrane
• highly permeable to water but
not to most of the electrolytes
• large amounts of K+ and PO43-
ions
• moderate quantities of Mg2+
and SO42- ions,
• large amounts of protein, ~4x
as much as in plasma
• small quantities of Na+ and Cl-
almost no Ca2+ ions.
MEASUREMENT OF FLUID VOLUMES IN THE DIFFERENT BODY FLUID
COMPARTMENTS—THE INDICATOR DILUTION PRINCIPLE
• The volume of a fluid compartment in the body can be measured by

• placing an indicator substance in the compartment
• allowing it to disperse evenly throughout the compartment’s fluid
• then analyzing the extent to which the substance becomes diluted
• “indicator-dilution” method of measuring the volume of a fluid compartment

• based on the principle of conservation of mass
• the total mass of a substance injected into a fluid compartment = the total mass after
dispersion in the compartment
ASSUMPTIONS
• 1) the indicator disperses evenly throughout the compartment.
• (2) the indicator disperses only in the compartment that is being measured.
• (3) the indicator is not metabolized
• A small, known amount of dye or other indicator
contained in a syringe injected into a chamber
• the substance is allowed to disperse throughout the
chamber until it becomes mixed in equal
concentrations in all areas.
• a sample of fluid containing the dispersed substance
is removed
• the concentration is analyzed chemically,
photoelectrically, or by other means
• Mass = volume x concentration
• Let’s call the syringe A and the chamber B
• Mass A = Mass B
• If none of the substance leaks out of the compartment,
the total mass of substance in the compartment
(Volume B × Concentration B) will equal the total mass
of the substance injected (Volume A × Concentration
A). Need to know (1) the total amount of substance
injected into the chamber and (2) the concentration
• By simple rearrangement of the equation, we can of the fluid in the chamber after the substance
calculate the unknown volume of chamber B has been dispersed.
E.g 1:
• A solution containing 5 g of Evans blue is injected into a chamber containing an
unknown volume of fluid. The fluid was mixed evenly and a small sample extracted to
determine the concentration of the fluid solution. The concentration was found to be
0.2M. Calculate the volume of the fluid into which the dye was injected.
• Indicator Mass A = Volume A x Concentration A
• Indicator Mass B = Volume B x Concentration B
• Indicator Mass A = Indicator Mass B
• Therefore we use the formula,
• Indicator Mass B = Volume B x Concentration B
• 5 g = Volume B x 0.2
• Volume B = 5 / 0.2 = 25L
(tritiated water)
SAMPLE PROBLEM
• A 65-kg man is participating in a research study for which it is necessary to
know the volumes of his body fluid compartments. To measure these
volumes, the man is injected with 100 mCi of D2O and 500 mg of mannitol.
During a 2-hour equilibration period, he excretes 10% of the D2O and 10% of
the mannitol in his urine. Following equilibration, the concentration of D2O in
plasma is 0.213 mCi/100 mL and the concentration of mannitol is 3.2 mg/100
mL.
• What are his total body water, his ECF volume, and his ICF volume?
• Is the man's total body water appropriate appropriate for his weight?
• Costanzo, Linda S.. Physiology E-Book (p. 250). Elsevier Health Sciences. Kindle Edition.
SOLUTION
• Total body water= Amount of D2O injected−Amount of D2O excreted
Concentration of D2O
• (100 mCi−(10% of 100 mCi)) / 0.213 mCi/100 mL
• 90 mCi/0.213 mCi/100 mL
• 90 mCi/2.13 mCi/L
• =42.3 L
• ECF volume = Amount of mannitol injected – Amount of mannitol excreted

Concentration of mannitol
• (500mg−(10% of 500 mg)) / 3.2 mg/100 mL
• 450 mg/3.2 mg/100 mL
• 450 mg/32 mg/L
• =14.1 L
• ICF volume=Total body water−ECF volume
• 42.3 L−14.1 L=28.2 L
SOLUTION
• The man's total body water is 42.3 L, which is 65.1% of his body weight (42.3 L is
approximately 42.3 kg; 42.3 kg/65 kg = 65.1%). This percentage falls within the normal
range of 50%–70% of body weight.
TERMINOLOGY
• Solution: any homogenous mixture; most frequently in liquid state
• Solute: the substance whose physical state is preserved when a solution is formed
• Percent solution: the concentration of solute in grams per 100mL of aqueous solution.
e.g A 5% aqueous solution of dextrose would contain 5g of dextrose and sufficient
water to make 100mL of solution.
• Specific gravity of solutions: the number of times heavier an object is than an equal volume
of water at same temperature ; no units
• Specific gravity of blood - the ratio of weight of a volume of blood to the weight of same
volume of water (approx. 1.06)
• Specific gravity of urine - the ratio of weight of a volume of urine to the weight of same
volume of water (between 1.003 and 1.035)
• Selectively permeable membrane: a membrane that permits the passage not only of solvent
but also of selected solutes (e.g most cell membranes in the body).
• Impermeant solute: solute that cannot cross the cell membrane
OSMOLARITY, OSMOTIC PRESSURE
AND CHANGES IN VOLUME OF THE
BODY FLUID COMPARTMENTS
BASIC PRINCIPLES OF OSMOSIS
• Osmosis is the net diffusion of water Selectively
across a selectively permeable Permeable
membrane from a region of high membrane
water concentration to one that has a
lower water concentration.
• When an impermeant solute is added
to pure water→ the solute
concentration increases → water
concentration decreases.
• Water diffuses from a region of low
solute concentration (high water
concentration) to one with a high
solute concentration (low water water
concentration).
solute
OSMOSIS ACROSS CELL MEMBRANES AND
REGULATION OF FLUID EXCHANGE
• Cell membranes are

Extracellular
relatively impermeable to Cell
most solutes but highly membrane
permeable to water (i.e.
selectively permeable) Intracellular
• Thus, if a solute such as
sodium chloride is added
to the extracellular fluid,
water rapidly diffuses from
the cells through the cell
membranes into the
extracellular fluid until the
water concentration on
both sides of the
membrane becomes
equal.
OSMOSIS ACROSS CELL MEMBRANES AND
REGULATION OF FLUID EXCHANGE
• Conversely, if a solute such as Extracellular

sodium chloride is removed
Cell
from the extracellular fluid, membrane
water diffuses from the Intracellular
extracellular fluid through the
cell membranes and into the
cells.
• The distribution of fluid

between intracellular and
extracellular compartments is
determined mainly by the
osmotic effect of the smaller
solutes— especially sodium,
chloride, and other
electrolytes— acting across
the cell membrane.
TERMS RELATED TO SOLUTES/ OSMOSIS IN
BODY FLUIDS
• Moles
• Molarity
• Equivalents
• Osmolarity
• Osmolality
• Osmotic Pressure
• Tonicity
RECALL: MOLES
1 mole = 6.02 × 1023 of particles
• 1 mol of C = 6.02 × 1023 carbon atoms
• 1 mol of H2O = 6.02 × 1023 water molecules
1 mole = 1000mmol = 1000000µ

µmol
1 mole = the sum of the atomic masses (g)

1 mole of CaCl2 = 111.1 g
MW of Ca = 40.1 g/mol
MW of Cl = 35.5 g/mol
40 + 35.5(2) = 111.1 g/mol CaCl2
1 mol per Litre = 1 Molar (1M)

EQUIVALENTS
• The equivalent (symbol: Eq ), sometimes termed the molar equivalent

• A unit of electrical charge used in chemistry and the biological sciences
• The equivalent of substance A = amount of substance A (mols) multiplied by its valence
• 1 mmol of Na+ is equal 1 mEq
• 1 mmol of Ca2+ is equal 2 mEqs
• Normally expressed as milliequivalents of solute per litre of solvent

• or milliNormal, where mEq/L = mN
• This is especially common for measurement of compounds in biological fluids
• for instance, the healthy level of potassium in the blood of a human is defined between
3.5 and 5.0 mEq/L.
OSMOLES
• Because the water concentration of a solution depends on the number of solute particles in the
solution, a concentration term is needed to describe the total concentration of solute particles,
regardless of their exact composition.
• The total number of particles in a solution is measured in osmoles.
• One osmole (osm) is equal to 1 mole (mol) (6.02 × 1023) of solute particles.
• A solution containing 1 mole of glucose (which is covalent and therefore does not
dissociate) in each liter has a concentration of 1 osm/L.
• A solution that contains 1 mole of a molecule that dissociates into two ions, such as
sodium chloride (NaCl) will contain 2 osm/L.
• A solution that contains 1 mole of a molecule that dissociates into three ions, such as
sodium sulfate (Na2SO4), will contain 3 osm/L.
• Thus, the term osmole refers to the number of osmotically active particles in a solution rather
than to the molar concentration.
• In general, the osmole is too large a unit for expressing osmotic activity of solutes in the body
fluids. The term milliosmole (mOsm), which equals 1/1000 osmole, is commonly used.
CALCULATING OSMOLES
• One liter of solution contains a mixture of 6 g of NaCl (MW (58.5g) and 210mg of MgCl2 (MW
95.24g). What are the osmolarities of NaCl and MgCl2 in the solution?
STEP 1 – calculate the number of millimoles of NaCl and MgCl2 present in the solution
• 58.5g = 1 mol of NaCl
6 g = 1/58.5 x 6 = 0.102 mols/L = 0.102 M = 102mM NaCl
• 95.24 g = 95240mg = 1 mol of MgCl2
210mg = 0.0022M = 2.2mM MgCl2
STEP 2 – calculate the number of milliosmoles of NaCl and MgCl2 present in the solution
• NaCl = 102 x 2 ions/molecule = 204 mOsm
• MgCl2 = 2.2 x 3 ions/ molecule = 6.6 mOsm
OSMOLARITY AND OSMOLALITY
• Osmolarity is expressed as osmoles per liter of solution.
• The osmolality of a solution is the concentration expressed as osmoles per

kilogram of solution;
• In most cases, it is easier to express body fluid quantities in liters of fluid

rather than in kilograms of water.
OSMOTIC PRESSURE
• Osmosis of water molecules
across a selectively permeable Extracellular
membrane can be opposed by
applying a pressure in the Cell
direction opposite that of the
membrane
osmosis. Intracellular
• The precise amount of
pressure required to prevent
osmosis is called the osmotic
pressure.
• Osmotic pressure, therefore, is
an indirect measurement of the
water and solute
concentrations (osmolarity or
osmolality) of a solution.
• The higher the osmotic
pressure of a solution,
the lower the water
concentration and the
higher the solute
concentration of the
solution.
1 mole of NaCl 1 mole of MgCl2
Osmotic
pressure
1L of water 1L of water
RELATION BETWEEN OSMOTIC PRESSURE AND
OSMOLARITY
• The osmotic pressure of a solution is directly proportional to the concentration of osmotically

active particles in that solution (i.e its osmolarity).
A B C
Na+
Cl-
1 mol Albumin (1 osm/L) 1 mol Glucose (1 osm/L) 1 mol NaCl (2 osm/L)

MW 70,000 MW 180
Osmotic pressure of A = Osmotic pressure of B Osmotic pressure twice that of A or B

i.e the osmotic pressure depends on the num ber of solutes and not the weight or size of solutes
OSMOTIC PRESSURE FORMULA
Π = i MRT
where
• i is the van 't Hoff factor (number of ion particles per individual molecule of solute, e.g. i =
2 for NaCl; 3 for BaCl2)
• M is the molarity
• R=0.08205746 L atm K-1 mol-1 is the gas constant
• T is the thermodynamic (absolute) temperature (Kelvins)
OSMOLARITY OF THE BODY FLUIDS
• The slight difference

between plasma and
interstitial fluid is caused
by the osmotic effects of
the plasma proteins,
which maintain about 20
mm Hg greater pressure
in the capillaries than in
the surrounding
interstitial spaces.
OSMOTIC EQUILIBRIUM IS NORMALLY MAINTAINED
BETWEEN INTRACELLULAR AND EXTRACELLULAR FLUIDS
• Large osmotic pressures can develop across the cell membrane with relatively small
changes in the concentrations of solutes in the extracellular fluid.
• However, larger changes in water or solute concentration can change the volumes of
intracellular and extracellular fluids
EXTRACELLULAR AND INTRACELLULAR FLUIDS IN
ABNORMAL STATES
• Some of the different factors that can cause extracellular and intracellular volumes to
change markedly are
• ingestion of water
• dehydration
• intravenous infusion of different types of solutions
• loss of large amounts of fluid from the gastrointestinal tract
• loss of abnormal amounts of fluid by sweating or through the kidneys
FLUIDS USED IN CLINICAL PRACTICE:
• Isotonic
• If a cell is placed in a solution with the same concentration of impermeant solutes
(i.e. having an osmolarity of 282 mOsm/L), the cells will not shrink or swell because
the water concentration in the intracellular and extracellular fluids is equal and the
solutes cannot enter or leave the cell.
• Isotonic solutions neither shrinks nor swells the cells.
• Examples: a 0.9 per cent solution of sodium chloride or a 5 per cent glucose
solution. These solutions are important in clinical medicine because they can be
infused into the blood without the danger of upsetting osmotic equilibrium between
the intracellular and extracellular fluids
• Hypotonic
• If a cell is placed into a hypotonic solution which has a lower concentration of
impermeant solutes (less than 282 mOsm/L), water will diffuse into the cell, causing
it to swell
• Water will continue to diffuse into the cell, diluting the intracellular fluid while also
concentrating the extracellular fluid until both solutions have about the same
osmolarity.
• Example: Solutions of sodium chloride with a concentration of less than 0.9 per cent
• Hypertonic Fluids
• If a cell is placed in a hypertonic solution having a higher concentration of
impermeant solutes, water will flow out of the cell into the extracellular fluid,
concentrating the intracellular fluid and diluting the extracellular fluid.
• In this case, the cell will shrink until the two concentrations become equal.
• Example: Sodium chloride solutions of greater than 0.9 per cent
EFFECT OF ADDING SALINE SOLUTION TO THE
EXTRACELLULAR FLUID
• If an isotonic saline solution is added to the extracellular fluid compartment,
• The osmolarity of the extracellular fluid does not change→ no osmosis occurs through the
cell membranes.
• Increase in extracellular fluid volume .The sodium and chloride largely remain in the
extracellular fluid because the cell membrane is virtually impermeable to sodium chloride.
• If a hypertonic solution is added to the extracellular fluid,
• The extracellular osmolarity increases and causes osmosis of water out of the cells into
the extracellular compartment.
• Increase in extracellular volume (greater than the volume of fluid added), a decrease in
intracellular volume
• If a hypotonic solution is added to the extracellular fluid,
• the osmolarity of the extracellular fluid decreases and some of the extracellular water
diffuses into the cells
• Both the intracellular and the extracellular volumes are increased by the addition of
hypotonic fluid, although the intracellular volume increases to a greater extent.
ISOSMOTIC, HYPEROSMOTIC, AND HYPO-
OSMOTIC FLUIDS
• The terms isotonic, hypotonic, and hypertonic refer to whether solutions will cause a change in cell volume.
• depends on the concentration of impermeant solutes. Some solutes, however, can permeate the cell
membrane.
• The terms iso-osmotic, hypo-osmotic and hyper-osmotic refer to relationships in osmolarity
• does not depend on whether solutes can penetrate the cell membrane.
• Iso-osmotic solutions have an osmolarity which is the same as the osmolarity within a cell (approx.
282mOsm/L)
• Hyper-osmotic and hypo-osmotic refer to solutions that have a higher (>282mOsm/L) or lower
osmolarity (< 282mOsm/L), respectively, than the osmolarity within a cell
• Some solutes are highly permeable across the cell membrane
• E.g urea,
• Such solutes can cause transient shifts in fluid volume between the intracellular and extracellular
fluids, but given enough time, the concentrations of these substances eventually become equal in the
two compartments and have little effect on intracellular volume.
GLUCOSE AND OTHER SOLUTIONS
ADMINISTERED FOR NUTRITIVE PURPOSES
• Many types of solutions are administered intravenously to provide nutrition to people who
cannot otherwise take adequate amounts of nutrition.
• Glucose solutions (widely used)
• Amino acid
• Homogenized fat solutions
• When these solutions are administered, they are:
• Isotonic
or
• They are given slowly enough that they do not upset the osmotic equilibrium of the body
fluids
• After the glucose or other nutrients are metabolized,
• an excess of water often remains
• kidneys excrete this in the form of a very diluted urine
FREEZING POINT DEPRESSION
• Freezing-point depression describes the process in which adding a solute to a solvent
decreases the freezing point of the solvent
• Examples include salt in water (seawater does not freeze at 0°C like pure water does )
∆TF = KF · m · i
Where,
• ∆TF, the freezing point depression, is defined as TF (pure solvent) - TF (solution).
• KF, the cryoscopic constant, which is dependent on the properties of the solvent, not the solute.
For water, KF = 1.853 °C·kg/mol.
• m is the molality (mol solute per kg of solvent)
• i is the van 't Hoff factor (number of ion particles per individual molecule of solute, e.g. i = 2 for
NaCl; 3 for BaCl2).
CALCULATIONS INVOLVING MOLECULAR
WEIGHT AND FREEZING POINT DEPRESSION OF
NON-ELECTROLYTE SUBSTANCES
Problem:
• The freezing point of a solution that is isotonic with blood contains 10.26 g of a non electrolyte
dissolved in 100mL (approx. 100g) of distilled water is found to be -0.56 oC. The freezing point
of pure water is 0 oC. The cryoscopic constant of pure water is 1.853 °C·kg/mol.
(i) What is the molecular weight of the non-electrolyte?
(ii) What is the osmotic pressure of the solution at 37oC.
Strategy:
(i)
Step 1: Calculate the freezing point depression of the solution
∆Tf = (Freezing point of pure solvent) - (Freezing point of solution)

(0 oC) - (-0.56 oC) = 0.56 oC
Step 2 : Calculate the molal concentration of the solution using the freezing point
depression
∆Tf = (Kf) (m) (i)

0.56 = 1.853 (m) (1)
m = (0.56 oC) / (1.853 oC/molal)
m = 0.3 molal
Step 3: Calculate the molecular weight of the unknown using the molal concentration
m = Number of grams of substance dissolved in 1000grams of water/ Mol. Wt

0.3 = 10.26 x 10/ Mol. Wt.
Mol. Wt. = 342g
(ii)
Π = i MRT,
where
• i is the van 't Hoff factor
• M is the molarity
• R=0.08205746 L atm K-1 mol-1 is the gas constant
• T is the thermodynamic (absolute) temperature (Kelvins)
Π = i MRT
= 0.3 x 0.082 x 310.15
= 7.63 Atmospheres 1 Atm = 760mmHg
ELECTROLYTE SUBSTANCES
• Problem: Calculate the mass of potassium chloride (mol. Wt. 74.6) in 100mL of distilled
water, which is isotonic with semen. The freezing point of semen is -0.6°C. The
cryoscopic constant is 1.853 kg/mol.
Strategy:
Step 1: Calculate the freezing point depression of the solution
∆Tf = (Freezing point of pure solvent) - (Freezing point of solution)

(0 oC) - (-0.6 oC) = 0.6 oC
ELECTROLYTE SUBSTANCES
Step 2 : Calculate the molal concentration of the solution using the freezing point depression
∆Tf = (Kf) (m) (i)

0.6 = 1.853 (m) (2)
m = 0.6/1.853(2) = 0.16 mol /kg water
Step 3: Calculate the mass of KCl using the molal concentration

0.16 = Number of grams of substance dissolved in 1000grams of water/ 74.6
Number of grams of substance dissolved in 1000grams of water is 11.94 g
Therefore, 1.194g must be weighed and dissolved in 100mL of water
PRACTICE QUESTIONS
Problem: Calculate the mass of glucose (mol. Wt. 180.16) in 100mL of distilled water,
which is isotonic with semen. The freezing point of semen is -0.6°C.
The cryoscopic constant is 1.853 kg/mol.
∆Tf = (Freezing point of pure solvent) - (Freezing

point of solution)
(0 oC) - (-0.6 oC) = 0.6 oC
∆Tf = (Kf) (m) (i)
0.6 = 1.853 (m) (1)
m = 0.6/1.853(1) = 0.32 mol /kg water

0.32 = Number of grams of substance dissolved in 1000grams of water/ 180.16
Number of grams of substance dissolved in 1000grams of water is 57.65 g
Therefore, 5.765 g must be weighed and dissolved in 100mL of water
PRACTICE QUESTIONS
Calculate the osmolarities of the following:
59g of KCl in 2 L (MW 74.55)
12657mg of MgSO4 in 3.5 L(MW 120.37)
7 mol of Ca(OH)2 in 5 L (MW 74.09)

PRACTICE QUESTIONS
Calculate the osmolarities of the following:
59g of KCl in 2 L (MW 74.55)

74.55 g = 1 mol
59g = 1/74.55 x 59
=0 .79mol in 2L
2L = 0.79 mol
1L = 0.79/2 = 0.396 mol/L
0.396 x 2 osm/L
0.792 Osm/L
PRACTICE QUESTIONS
12657mg of MgSO4 in 3.5 L(MW 120.37)

120.37 = 1 mol
120370 = 1 mol
12657 = 0.105 mol
0.105 mol in 3.5L

3.5 = 0.105
1 = 0.105/3.5 = 0.03 mol/L
0.03 x 2 osm/L
0.06 Osm/L
PRACTICE QUESTIONS
7 mol of Ca(OH)2 in 5 L (MW 74.09)
7mol = 5 L
1L =7/5
=1.4mol
1.4 x 3 = 4.2 Osm/L

PRACTICE QUESTIONS
Calculate the following concentrations in terms of millliequivalents:
5.5 mmol of K+
5.5 mEq
7 mmol of Al3+
21mEq
3.2 mmol of Mn2+

6.4 mEq
THE PH SCALE
• pH is defined as the negative logarithm

of the concentration of H+
STRONG ACIDS
• Strong acids and bases dissociate

completely in water
HCl + H2O Cl- + H3O+
• Cl- is the conjugate base of HCl
• H3O+ is the conjugate acid of H2O
ACETIC ACID IS A WEAK ACID
• Weak acids and bases do not dissociate

completely in H2O
THE HENDERSON-HASSELBALCH EQUATION
HA ⇔ H+ + A-
Ka = [H+] [A-]
[HA]
Rearranging the equation and using logs gives you:
THE HENDERSON-HASSELBALCH EQUATION
• Defines the pH of a solution in terms of:

(1) The pKa of the weak acid
(2) Concentrations of the weak acid
(HA) and conjugate base (A-)
BUFFERS
HA ⇔ H+ + A-
weak acid conjugate base
A good buffer : [HA] = [A-]

H+ + A- → HA
OH- + HA → H2O +A-
BUFFERED SOLUTIONS RESIST CHANGES IN PH
• Buffer capacity is the ability of a solution to resist

changes in pH
• Most effective buffering occurs where:
solution pH = buffer pKa
• At this point: [weak acid] = [conjugate base]
• Effective buffering range is usually at pH values
equal to the pKa ± 1 pH unit
BUFFERS IN LIVING ORGANISMS
• Intracellular buffers
• (HPO42-/H2PO4-) buffer
• Histidine buffer
• Extracellular buffers
• HCO3-/H2CO3
THE (HPO42-/H2PO4-) BUFFER
pH = pKa + log10([HPO42-]/[H2PO4-])
7.4 = 7.2 + log10 ([HPO42-]/[H2PO4-])
THE HISTIDINE BUFFER
• Imidazolium group is
Present in the amino acid
Histidine
• In histidine the pKa is 6.04
• Would this be a suitable buffer within cells
and living systems?
• But the histidine present in the
dipeptide anserine (which contains
alanine and histidine) has a pKa of
7.04
• Found in skeletal muscle and brain of
mammals and birds
THE (HCO3-/H2CO3) BUFFER
• Blood plasma of mammals has a constant pH

which is regulated by a buffer system of:
carbon dioxide /carbonic acid /bicarbonate
• Buffer capacity depends upon equilibria between:
(1) Gaseous CO2 (air spaces of the lungs)
(2) Aqueous CO2 (dissolved in the blood)
(3) Carbonic acid
(4) Bicarbonate
THE (HCO3-/H2CO3) BUFFER
FIG. 2.21
Regulation of the pH
of blood in mammals
REFERENCES
• Physiology of Body Fluids. Dr. Affan Ezzat. Lecture 3. www.comed.uobaghdad.edu.iq/
Cholesterol metabolism

Biochemistry unit
Objectives
At the end of this topic you should be able to:
• Outline the biochemical pathways involved

in the synthesis of cholesterol and its
derivatives
• Discuss the regulation of cholesterol
synthesis and turnover and the use of
enzyme inhibitors to modulate cholesterol
levels
Cholesterol metabolism
Cholesterol is a sterol, present in cell membrane, brain and
lipoprotein
• It is a precursor for all steroids
• About 1 g of cholesterol is synthesized per day in humans
• It is an amphipathic lipid
• Lipoproteins transports the free cholesterol in the circulation
• Cholesterol ester is a storage form of cholesterol found in most
tissues
• 80% of the liver cholesterol converted to bile acids
Vitamin D3 formed from 7-dehydrocholesterol.
• All the steroids have cyclopentanoperhydrophenanthrene
ring. Made up of three cyclohexane rings, A,B and C and a
cyclopentane ring D
• Normal Blood level is 150-200 mg%
• Hypercholesterolemia seen in nephrosis, diabetes

mellitus, hypothyroidism (may be due to reduced HDL
receptors on hepatocytes) and obstructive jaundice
• Increased cholesterol level leads to atherosclerosis
• The OH group in the 3rd position can get esterified to fatty

acids to form cholesterol esters. This esterification occurs in
the body by transfer of PUFA moiety by Lecithin cholesterol
acyl transferase. This step is important in the regulation of
cholesterol level.
• It is a poor conductor of electricity

SYNTHESIS
• Site: Extra Mitochondrial. The enzymes
involved are found in cytosol and
microsomal fractions of the cell.
• Synthesis takes place in liver, skin and
intestine and also in adrenal cortex &
testis.
• All the 27 carbon atoms are derived
from acetyl CoA
• Total of 18 acetyl Co A are required
• Acetyl CoA formed in glycolysis and -
Oxidation of fatty acid are the
precursors for the cholesterol synthesis
Regulation of Cholesterol synthesis
 Cholesterol biosynthesis is controlled by the rate limiting enzyme

HMG-CO A reductase
1. Feedback control: The end product cholesterol controls its own

synthesis of the enzyme by a feedback mechanism. Increase in the
cellular concentration of cholesterol reduces the synthesis of the
enzyme by decreasing the transcription of the gene responsible for
the production of HMG CoA reductase.
2. Hormonal regulation: The HMG CoA reductase exists in two

interconvertible forms.
Insulin and thyroid hormones Increase HMG CoA reductase activity
The dephosphorylated form of the enzyme is more active,
phosphorylated is less active. Hormones exert their influence through
cAMP
 Glucagon and glucocorticoids decrease HMG-CoA
reductase activity
 Inhibition by drugs: The drugs Compactin and

lovastatin, mevastatin, simvastin are competitive
inhibitors used to decrease the cholesterol.
 HMG CoA reductase is inhibited by bile acids.
 LDL transports cholesterol from the liver to

peripheral tissues.
 HDL transports cholesterol from tissues to liver

Compactin, lovastatin [Competitive inhibitors]
Mevastin, Simvastin
HMG CoA _
_
Insulin, thyroxin + HMG CoA Reductase Glucagon
(dephosphorylates enz) glucocorticoids
(Phosphorylates enz)
Mevalonate Translation
mRNA
Cholesterol _ Transcription
DNA
 Glucagon and glucocorticoids inactivate the enzyme through

phosphorylation
 Insulin, thyroxin activate the enzyme through dephosphorylation
METABOLIC FATE OF CHOLESTEROL
Cholesterol is converted into following compounds as shown below.

Cholesterol is mainly excreted in the form of bile salts in stool.
Steroid hormone
(Testosterone, estrogens
Acetyl CoA Cholesterol progesterone,glucocorticoids
mineralocorticoids)
Vitamin D3
Bile acids [salts]
Increased plasma cholesterol results in the accumulation of cholesterol
under the tunica intima of the arteries causing atherosclerosis. The
progression of the disease process leads to narrowing of the blood vessels.
Dietary intake of polyunsaturated fatty acid (PUFA) helps in transport and
metabolism of cholesterol and prevents atherosclerosis
Role of LCAT:
High density lipoprotein (HDL) and the enzyme lecithin-cholesterol

acyl transferase (LCAT) are responsible for the transport and
elimination of cholesterol from the body.
LCAT is a plasma enzyme, synthesized by the liver.
LCAT catalyses the transfer of fatty acid from the second position of
phosphatidyl choline (lecithin) to the OH group of cholesterol.
HDL cholesterol is the real substrate for LCAT and this reaction is
freely reversible.
LCAT activity is associated with apo-A1 of HDL.
Ref: Essentials of Biochemistry, Prof. S. Nayak

March 2020
DE NOVO SYNTHESIS OF FATTY ACID

Biochemistry unit
Objectives
At the end of this lecture the learner should

be able to explain
• The synthesis of fatty acids

• The regulation fatty acid synthesis in a
well fed and fasting state
BIOSYNTHESIS or DE NOVO SYNTHESIS OF FATTY ACID
The majority of the fatty acids required supplied through our diet.
Fatty acids are synthesised whenever there is a caloric excess in the our
diet.
The excess carbohydrate and protein obtained through diet can be
converted to fatty acids which are stored as triacylglycerol.
• Fatty acid synthesis involves the similar steps

involved in -oxidation of fatty acid but in a reverse
way.
• Mammals can synthesise major portion of the

saturated fatty acid as well as monounsaturated fatty
acids.
• The system for the fresh synthesis of fatty acid is

known as de novo synthesis of fatty acid
• Takes place in liver and lactating mammary glands
and to a lesser extent in adipose tissue, kidney .
• The enzyme machinery is located in cytoplasm.
• Enzyme system is referred to as extra mitochondrial or

cytoplasmic fatty acid synthase system.
• Palmitic acid is the major fatty acid synthesised
• All the 16 carbon atoms are from acetyl CoA.
• The acetyl CoA used as a primer and which forms

carbons 15 and 16 of palmitate. The addition of all
the subsequent 2-C units is through malonyl CoA
formation
• Acetyl CoA and NADPH are the prerequisites for the
fatty acid synthesis.
• Acetyl CoA produced in the mitochondria cannot

enter into cytoplasm through inner mitochondrial
membrane.
Therefore it combines with Oxaloacetate in
mitochondria to form citrate.
• Citrate is freely transported to cytosol where it is

cleaved by citrate lyase to liberate acetyl CoA and
Oxaloacetate.
Mitochondria Cytoplasm Glucose
Pyruvate Pyruvate HMP Shunt
NADPH+H+, CO2
Malic Enzyme NADP+
PDH Malate
NAD+
Malate Dh
FA, amino acids NADH+H+ FA
Oxaloacetate
Acetyl CoA + Oxaloacetate AcetylCoA, ADP+Pi

Citrate synthase Citrate lyase
CoASH, ATP
Citrate Citrate
For FA synthesis, 8 acetyl CoA are transported from the mitochondria

to cytosol, which is linked with the synthesis of 8 NADPH.
14 NADPH are required to synthesise one molecule of Palmitate.
The remaining 6 NADPH supplied from HMP shunt.
*Acetyl CoA carboxylase

1) Acetyl CoA malonyl CoA
Biotin, ATP, CO2 ADP+Pi
* Regulatory enzyme in FA synthesis.
• The remaining reactions of FA synthesis are catalysed by

multifunctional enzyme known as fatty acid synthase complex [FAS]
• FAS is a dimer with two identical subunits.
• Each monomer possesses the activities of seven different enzymes
and an acyl carrier protein (ACP) bound to 4’phosphopantetheine-SH
group.
• Two subunits lie in antiparallel (head to tail) orientation.
• The-SH group of phosphopantetheine of one subunit is in close
proximity to the –SH of cysteine residue of the other subunit.
• Each monomer of FAS contains all the enzyme activities of fatty
acid synthesis.
• Dimer form of enzyme is functionally active. [functional unit
consists of half of each subunit interacting with the complimentary
half of the other].
Components of fatty acid synthase complex:

1. Acetyl transferase [AT]
2. Malonyl transferase [MT]
3. -Keto acyl synthase [KS]
4. -Keto acyl reductase [KR]
5. -Hydroxy acyl dehydratase [HD]
6. Enoyl reductase [ER]
7. Thioestarase [TE]
Acyl carrier protein [ACP].
Functional division
KS AT MT HD ER KR ACP TE
Cys 4’ phosphopantetheine
SH SH
-----------------------------------------------------------------------Subunit
SH SH division
4’ phosphopantetheine Cys
TE ACP KR ER HD MT AT KS
1. Fatty acid synthesis starts with the transfer of an acetyl CoA
to cysteinyl SH group of ACP
Acetyl CoA + (CE)-SH AT Acetyl S-(CE) + CoA --1
2. Malonyl CoA-ACP transferase transfers malonate from

malonyl CoA to bind to ACP
Malonyl CoA+ACP-SH MT Malonyl-S-ACP + CoA—2
3) The acetyl unit attached to cysteine is transferred to

malonyl group attached to ACP. Malonyl moiety loses
CO2,which was added by acetyl CoA carboxylase & form
β-ketoacyl enzyme.
4) -Ketoacyl-enzyme is reduced to -hydroxy butyryl
enzyme complex using NADPH+H+.
5) Molecule of H2O is removed from -OH butyryl enzyme

to form ,  unsaturated acyl enzyme.
6) The unsaturated bond in ,  unsaturated acyl enzyme

is again reduced using NADPH+H+ to form butyryl or
acyl enzyme.The carbon chain attached to ACP is
transferred to cysteine residue and the reactions 2-6
are repeated 6 more times and finally palmitic acid is
synthesised.
7) The completely synthesized fatty acid is released from

the enzyme system by the action of thioesterase enzyme.
Fatty acid chain elongation and desaturation occurs in the
microsomes of endoplasmic reticulum and mitochondria.
Of the 16 carbons present in palmitate, only two come from acetyl CoA directly.
The remaining 14 are from malonyl CoA (produced from acetyl CoA).
During elongation in microsomes palmitate activated to palmitoyl CoA.

Malonyl Co serves as the donor of two carbons at a time in series of reactions.
Major elongation reaction occurs in the body involves the formation of stearyl
CoA [C18] from palmitoyl CoA [C16]
Elongation of this stearyl CoA in brain increases during myelination to provide
C22 and C24 fatty acids of sphingolipids
Mitochondrial elongation is less active and uses acetyl CoA as the source of two
carbon units
8AcetylCoA+7ATP+14NADPH+14H+Palmitate+8CoA+7ADP+7Pi+6H2O+14NADP+
Regulation
Acetyl CoA carboxylase enzyme controls a committed step in fatty acid

synthesis. This enzyme exists as an inactive monomer or an active
polymer. Citrate promotes polymer formation, hence increases FA
synthesis. Palmitoyl CoA causes depolymerisation of the enzyme
and inhibit FA synthesis.
Hormonal influence
• Glucagon, epinephrine & norepinephrine inactivate the enzyme by

cAMP dependent phosphorylation and inhibits FA synthesis
• Insulin dephosphorylates & activates the enzyme and promotes FA
synthesis.
Dietary Regulation:
• High carbohydrate and fat free diet increases the

synthesis of Acetyl CoA carboxylase and FA
synthase,which promotes FA synthesis.
• Fasting and high fat diet decreases FA production.

NADPH influences FA synthesis.
Dr. S. Nayak March 2020

Digestion

EWMSC, Mount Hope
Trinidad
Objectives
At the end of this course student should be able to
• Describe digestion and absorption of the constituents of

fats, carbohydrates and proteins
• List the enzymes of gastrointestinal tract
Dr.S. Nayak 2
Digestion
 Diet contains carbohydrates, fat and proteins
 They are high molecular weight complex compounds.
 Must be digested for absorption
Digestion of carbohydrates
• The major carbohydrates of our diet are Polysaccharides (starch
and glycogen)
• Hydrolyzed to simple sugars through enzymes.
• Salivary amylase hydrolyses -1,4-glycosidic linkages of
polysaccharide chain to produce mono and disaccharides.
• Further digestion happens in the small intestine by the
intestinal enzymes (which hydrolyze terminal -1,4-glycosidic
linkage).
Dr.S. Nayak 3
•Entry of acidic contents of stomach into duodenum, stimulate the
mucosal cells to release secretin and cholecystokinin
•Secretin stimulates the pancreas to release bicarbonate and water

to neutralize the acidic chyme from the stomach
•Cholecystokinin stimulates the production of digestive enzymes

including pancreatic amylase
•Pancreatic amylase digest the polysaccharides to maltose,

isomaltose and a limit dextrin.
•Disaccharidases such as maltase, lactase and sucrase digest

disaccharides like maltose, lactose and sucrose respectively into
their respective monosaccharide units.
Dr.S. Nayak 4
Dr.S. Nayak 5
Dr.S. Nayak 6
•Humans does not produce β 1,4-endoglucosidase in
digestive juice to digest cellulose.
•Cellulose helps in easy peristalsis and provides bulk to the
feces.
•Lactase deficiency leads to lactose intolerance.
Absorption of monosaccharides
•The galactose and glucose are absorbed very rapidly by the
active process
•Fructose and mannose are absorbed by a Na+ independent

facilitative transport mechanism
Dr.S. Nayak 7
Dr.S. Nayak 8
Digestion of proteins
Stomach:
•Protein does not undergo digestion in the mouth.
•Enters the stomach and stimulates the secretion of the
hormone gastrin (from gastric mucosal cells).
•Gastrin stimulates the release of gastric juice containing
HCl and pepsinogen [rennin in infants].
•The HCl is secreted by parietal cells, which unfolds the
proteins and activates the proteolytic enzyme pepsin.
Dr.S. Nayak 9
•Pepsin secreted by chief cells as pepsinogen & converted
to pepsin by HCl (zymogen activation).
•Pepsin digests protein polypeptides into tripeptides,
dipeptides and amino acids.
•Pepsin specifically hydrolyses the peptide bonds of protein
involving the aromatic amino acids or acidic amino acids
•Rennin in infants is also called as chymosin or rennet.
Intestine:
•Entry of acidic contents from the stomach into the
intestine triggers the secretion of the hormones such as
cholecystokinin and secretin.
Dr.S. Nayak 10
•Secretin stimulates the release of bicarbonate and
pancreatic juice from pancreas into the small intestine
•Cholecystokinin stimulates the secretion of pancreatic endo

and exopeptidases
•Endopeptidases cleave internal peptide bonds of proteins

to convert into smaller peptides
•The endopeptidases: trypsin, chymotrypsin, elastase, which

are secreted in pro-enzyme forms and are converted to
active forms by enteropeptidase (enterokinase)
Dr.S. Nayak 11
•Trypsin hydrolyses peptide bonds whose carboxyl groups are
contributed by lysine and arginine
•Chymotrypsin specifically hydrolyses the peptide bonds

involving the carboxyl group aromatic amino acids
(Phenylalanine, tyrosine or tryptophan). It also splits peptide
bonds of leucine, methionine asparagine and histidine
•Elastase hydrolyses those peptide bonds formed by non-

polar amino acids, such as alanine, serine and glycine
Dr.S. Nayak 12
Exopeptidases:
Carboxypeptidase and Aminopeptidase
•Carboxypeptidase is a composition of pancreatic juice
and hydrolyses the first peptide bond from the free
carboxyl end
•Aminopeptidase hydrolyses the first peptide bond from
the free amino terminal end
Absorption
•The absorption of amino acids includes Na+ dependent
active transport mechanism, which requires ATP as
energy source.
Dr.S. Nayak 13
DIGESTION AND ABSORPTION OF LIPIDS
•Digestion of lipids depends on the bile salts for the

emulsification.
•Lipids inhibit gastric motility and so retard the evacuation

of the stomach
•The gastric lipase and lingual lipase of chyme are active

only at neutral pH
•In adults no digestion takes place in stomach due to acidic

pH
Dr.S. Nayak 14
•The hydrophilic short and medium chain fatty acids are
absorbed via mucosal cell and enter the portal vein.
•The longer chain fatty acids dissolve in the diet and pass
into the duodenum
•Entry of acidic chyme from the stomach into the

duodenum stimulates the secretion of enteric hormones
like secretin and cholecystokinin by the mucosal cells of
duodenum
•Cholecystokinin
a) Helps in the contraction of gallbladder to release bile salts into the small
intestine
b) Acts on the exocrine cells of the pancreas, causing them to release

digestive enzymes including lipase
c) Decreases gastric motility, which results in a slow release of the gastric

contents into small intestine.
Dr.S. Nayak 16
•Secretin causes the pancreas to release a bicarbonate
rich solution which neutralizes the acidic chyme and
changes the pH to the alkaline side
•Formation of alkaline pH of the content is very

important for the action of lipase and intestinal enzymes
•Bile enters the duodenum and provides the emulsifying

action. After emulsification, the lipolytic enzymes such
as lipase, phospholipase A2 and cholesterol esterase of
pancreatic juice hydrolyse lipids.
Dr.S. Nayak 17
•Dietary glycerophospholipds are digested by pancreatic
phospholipase-A2 (hydrolysis of fatty acid residues at the 2nd
position of the glycerophospholipid to form lysophospholipid).
•Inside the mucosal cells fats (TG) are re-synthesized and

converted to chylomicron and transported to blood via lymphatic
vessel
•Fatty acids less than 10 carbon atoms along with glycerol are
carried by portal blood to the liver.
Dr.S. Nayak 18
Dr.S. Nayak 19
•Long chain free fatty acids, free cholesterol, 2-monoglyceride and 1-
monoglyceride and lysophospholipid together with bile salts form
mixed micelles.
•Bile salts aggregate with their hydrophobic region placed internally

and hydrophilic region facing the water medium and makes the
micelle water soluble. The glycerides and long chain fatty acids in
these micelles are transported into the intestinal mucosal cells
leaving bile salts in the medium itself. The bile salts are reabsorbed
in the intestine and returned to the liver by the portal vein for re-
secretion into the bile. This process is called enterohepatic
circulation of bile salts
Dr.S. Nayak 20
Dr.S. Nayak 21
•The 1-monoacylglycerol are further hydrolysed in the
intestinal mucosal cell by intestinal lipase
•The 2-monoacylglycerol are reconverted to triglyceride.

The utilization of fatty acids inside the mucosal cell for
the re-synthesis of triacylglycerol needs the activation to
acyl-CoA by thiokinase enzyme
•The absorbed lysophospholipids and cholesterol are also

recycled with acyl-CoA to regenerate phospholipids and
cholesterol esters.
Dr.S. Nayak 22
•The triacylglycerol, phospholipid, cholesterol ester
synthesized in the intestinal mucosal cell and the absorbed
fat soluble vitamins are transported from the mucosal cells
into the lymph in the form of chylomicron
•The absorbed lipids are either oxidized mainly in the liver

or stored in the depots (adipose tissue)
•For utilization by the body, triglycerides are first hydrolysed

by lipase to release glycerol and free fatty acids
•Glycerol is converted to glucose by gluconeogenesis or

enters into glycolysis. Fatty acids are oxidized to CO2 and
H2O with the liberation of large amount of energy.
Dr.S. Nayak 23
Vitamins and Minerals
DUODENUM: Fe, Ca, Mg, Zn
JEJUNUM: Vit C, B1 B2, B6, Folic Acid
ILEUM: Fat soluble vitamins are absorbed via micelles; B12
COLON: Na; K; vit K from bacterial action
ABSORPTION OF MINERALS
PHASE 1: INTRALUMINAL
•Chemical reactions and interactions in the stomach and
intestines
•Cations (eg. Ca 2+): influenced by pH of luminal contents
and composition of chyme from stomach
•Soluble in acid pH of stomach but form insoluble
hydroxides in the higher pH of the small intestine
•Kept available for absorption by ligands such as amino
acids, organic acids, sugars Dr.S. Nayak 24
PHASE 2: TRANSLOCATION
•Passage across the membrane into the intestinal mucosal cell
•Small anions: via simple diffusion
•Cations: facilitated diffusion or active transport . Often more than
one mechanism is available depending on concentration of trace
element
PHASE 3: MOBILIZATION
•Transport across the serosal surfaces of cell into blood or
Sequestered within the cell or BOTH
•Fe and Zn are either bound to proteins within the cell or added to
the intracellular pool.
•Bound forms can be added to the pool or remain bound and are lost
via desquamation
Reference: Essentials of Biochemistry by Dr S Nayak

Dr.S. Nayak 25
Gluconeogenesis and Cori cycle
Objectives
The student should be able to;
1.Describe gluconeogenesis
2.Explain how the gluconeogenesis regulated
3.Discuss the Cori cycle and its regulation
Prof S Nayak 1
Gluconeogenesis
• Mainly occurs in cytosol

• Some precursors are produced in mitochondria
• Takes place in liver and kidney
• Synthesis of glucose or glycogen from non carbohydrates
like pyruvate, lactate glucogenic amino acids, glycerol and
propionic acid
• Involves steps of TCA cycle and reversal of glycolysis
• The irreversible steps of glycolysis are catalysed by
Hexokinase
Phosphofructokinase and
Pyruvate kinase
• These three stages bypassed by alternate enzyme specific
to gluconeogenesis
• They are called as key enzymes of gluconeogenesis
Prof S Nayak 2
1. Pyruvate carboxylase
2. Phosphoenolpyruvate carboxykinase (PEPCK)
3. Fructose 1,6- Bis phosphatase
4. Glucose 6-phosphatase
The pathway meet the needs of the body for glucose
Continuous supply of glucose required as a source

of energy for the CNS, Brain, RBC and skeletal
muscle during starvation
Prof S Nayak 3
Prof S Nayak 4
Prof S Nayak 5
Substrates for Gluconeogenesis
Gly, Ala, Ser, Thr, Cys, Trp  pyruvate oxaloacetate
Phe, Tyr, fumarate

Asp, Asn
Arg, His,Glu, gln, proKGoxaloacetateGlucose
Val, isoleucine, Met  succinyl CoA
Propionyl CoA  succinyl CoA  oxaloacetate
Prof S Nayak 6
Prof S Nayak 7
Cori’s Cycle
Glucose/Glycogen converted to lactate in the muscle and
this lactate is converted back to glucose in liver
During active muscle contraction glycogen breaks down

to generate glucose
Prof S Nayak 8
Prof S Nayak 9
Regulation of gluconeogenesis
The glucagon and the availability of substrates mainly regulate

gluconeogenesis
Glucagon & glucocorticoid stimulate gluconeogenesis.
Insulin inhibit gluconeogenesis
Glucogenic amino acids have stimulating effect on key

gluconeogenic enzymes
•Acetyl CoA promotes gluconeogenesis
Starvationexcessive lipolysis in adipose

tissues acetyl CoA accumulates in the liver, acetyl CoA stimulate
gluconeogenic enzymes.
Prof S Nayak 10
Prof S Nayak 11
HEALTH TOPICS ▼   ≡
Glycemic index and glycemic load

for 100+ foods
Glycemic index and glycemic load offer information about how foods affect blood sugar and insulin. The lower a
food’s glycemic index or glycemic load, the less it affects blood sugar and insulin levels. Here is a list of the
glycemic index and glycemic load for more than 100 common foods.
FOOD Glycemic index Serving Glycemic load

(glucose = 100) size per serving
(grams)
BAKERY PRODUCTS AND BREADS
Banana cake, made with sugar 47 60 14
Banana cake, made without sugar 55 60 12
Sponge cake, plain 46 63 17
Vanilla cake made from packet mix with 42 111 24

vanilla frosting (Betty Crocker)
Apple, made with sugar 44 60 13
Apple, made without sugar 48 60 9
Waffles, Aunt Jemima® (Quaker Oats) 76 35 10
Bagel, white, frozen 72 70 25
Baguette, white, plain 95 30 15
Coarse barley bread, 75-80% kernels, 34 30 7

average
Hamburger bun 61 30 9
Kaiser roll 73 30 12
Pumpernickel bread 56 30 7
50% cracked wheat kernel bread 58 30 12
White wheat flour bread 71 30 10
Wonderâ¢ bread, average 73 30 10
Whole wheat bread, average 71 30 9
100% Whole Grain® bread (Natural Ovens) 51 30 7
Pita bread, white 68 30 10
Corn tortilla 52 50 12
Wheat tortilla 30 50 8
BEVERAGES
Coca Cola®, average 63 250 mL 16
Fanta®, orange soft drink 68 250 mL 23
Lucozade®, original (sparkling glucose 95Â±10 250 mL 40

drink)
Apple juice, unsweetened, average 44 250 mL 30
Cranberry juice cocktail (Ocean Spray®) 68 250 mL 24
Gatorade 78 250 mL 12
Orange juice, unsweetened 50 250 mL 12
Tomato juice, canned 38 250 mL 4
BREAKFAST CEREALS AND RELATED

PRODUCTS
All-Bran®, average 55 30 12
Coco Pops®, average 77 30 20
Cornflakes®, average 93 30 23
Cream of Wheat® (Nabisco) 66 250 17

Cream of Wheat®, Instant (Nabisco) 74 250 22
Grapenuts, average 75 30 16
Muesli, average 66 30 16
Oatmeal, average 55 250 13
Instant oatmeal, average 83 250 30
Puffed wheat, average 80 30 17
Raisin Bran® (Kellogg’s) 61 30 12
Special K® (Kellogg’s) 69 30 14
GRAINS
Pearled barley, average 28 150 12
Sweet corn on the cob, average 60 150 20
Couscous, average 65 150 9
Quinoa 53 150 13
White rice, average 89 150 43
Quick cooking white basmati 67 150 28
Brown rice, average 50 150 16
Converted, white rice (Uncle Ben’s®) 38 150 14
Whole wheat kernels, average 30 50 11
Bulgur, average 48 150 12
COOKIES AND CRACKERS
Graham crackers 74 25 14
Vanilla wafers 77 25 14
Shortbread 64 25 10
Rice cakes, average 82 25 17
Rye crisps, average 64 25 11

Soda crackers 74 25 12
DAIRY PRODUCTS AND ALTERNATIVES
Ice cream, regular 57 50 6
Ice cream, premium 38 50 3
Milk, full fat 41 250mL 5
Milk, skim 32 250 mL 4
Reduced-fat yogurt with fruit, average 33 200 11
FRUITS
Apple, average 39 120 6
Banana, ripe 62 120 16
Dates, dried 42 60 18
Grapefruit 25 120 3
Grapes, average 59 120 11
Orange, average 40 120 4
Peach, average 42 120 5
Peach, canned in light syrup 40 120 5
Pear, average 38 120 4
Pear, canned in pear juice 43 120 5
Prunes, pitted 29 60 10
Raisins 64 60 28
Watermelon 72 120 4
BEANS AND NUTS
Baked beans, average 40 150 6
Blackeye peas, average 33 150 10
Black beans 30 150 7

Chickpeas, average 10 150 3
Chickpeas, canned in brine 38 150 9
Navy beans, average 31 150 9
Kidney beans, average 29 150 7
Lentils, average 29 150 5
Soy beans, average 15 150 1
Cashews, salted 27 50 3
Peanuts, average 7 50 0
PASTA and NOODLES
Fettucini, average 32 180 15
Macaroni, average 47 180 23
Macaroni and Cheese (Kraft) 64 180 32
Spaghetti, white, boiled, average 46 180 22
Spaghetti, white, boiled 20 min, average 58 180 26
Spaghetti, wholemeal, boiled, average 42 180 17
SNACK FOODS
Corn chips, plain, salted, average 42 50 11
Fruit Roll-Ups® 99 30 24
M & M’s®, peanut 33 30 6
Microwave popcorn, plain, average 55 20 6
Potato chips, average 51 50 12
Pretzels, oven-baked 83 30 16
Snickers Bar® 51 60 18
VEGETABLES
Green peas, average 51 80 4
Carrots, average 35 80 2
Parsnips 52 80 4
Baked russet potato, average 111 150 33
Boiled white potato, average 82 150 21
Instant mashed potato, average 87 150 17
Sweet potato, average 70 150 22
Yam, average 54 150 20
MISCELLANEOUS
Hummus (chickpea salad dip) 6 30 0
Chicken nuggets, frozen, reheated in 46 100 7

microwave oven 5 min
Pizza, plain baked dough, served with 80 100 22

parmesan cheese and tomato sauce
Pizza, Super Supreme (Pizza Hut) 36 100 9
Honey, average 61 25 12
The complete list of the glycemic index and glycemic load for more than 1,000 foods can be found in the article
“International tables of glycemic index and glycemic load values: 2008” by Fiona S. Atkinson, Kaye Foster-Powell,
and Jennie C. Brand-Miller in the December 2008 issue of Diabetes Care
{http://care.diabetesjournals.org/content/31/12/2281.full}, Vol. 31, number 12, pages 2281-2283.
February 3, 2015
Updated: August 27, 2015
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Glycogen metabolism

St. Augustine
Trinidad
Objectives
At the end of this topic the student should be able to:
Describe the breakdown and the synthesis of glycogen.
Explain how the glycogen metabolism is regulated.
Describe the biochemical basis, signs and symptoms of glycogen storage diseases.
DR S NAYAK
2
Glycogen metabolism
· It is the storage from of glucose in animals

· Stored in liver (6-8%) and muscle (1-2%)
• Helps to maintain the blood glucose levels between
meals
• Glycogen stores increase in a well-fed state depleted
during fasting
· Muscle glycogen serves as a fuel reserve for the

supply of ATP during muscle contraction
In homopolysaccharide, glucose molecules held

together by - 1,4 linkages. Branch with -1, 6 linkage.
Glucokinase in liver and hexokinase in muscle which
converts glucose to glucose–6 phosphate
DR S NAYAK 3
Glycogenesis
 Synthesis of glycogen from glucose

 Occurs in liver and muscle
 Storage from in liver and muscle
 After the meal excess glucose is converted into
glycogen
 UDPG is the carrier of glucose
 Glucose from UDPG is attached at the non-reducing
end of glucose molecules of glycogen primer
DR S NAYAK 4
ATP ADP Phosphoglucomutase
Glucose Glucose 6-P Glu 1-P
Hexokinase
Glucokinase UDPG pyrophosphorylase
UTP
PPi
Uridine diphosphate glucose (UDPG)
Glycogen synthase
Glycogen primer UDP
Glycogen (1, 4 glucosyl units) n

(1, 4 and 1,6 Branching enzyme
Glucosyl units) n (Amylo-1, 4-1,6- transglucosidase)
(Glucosyl -4,6 transferase)
DR S NAYAK 5
Glycogenesis
DR S NAYAK 6
 In the absence of glycogen primer, GLYCOGENIN
(protein) can accept glucose from UDPG.
The initial glucose is attached to the OH group of
tyrosine residue of glycogenin.
The enzyme glycogen initiator synthase transfers the first
molecule of glucose to glycogenin.
Later glycogenin itself takes up a few glucose residue
to form a fragment of primer
 Branching enzyme (Amylo 1,4 –1,6 transglucosidase

transfers 6 glucose residues portion from one chain
to a neighbouring chain to form a -1,6 – linkage
DR S NAYAK 7
Glycogenolysis
Breakdown of glycogen to glucose
·Occurs in liver and muscle
·End product of liver glycogenolysis is glucose
·Muscle glycogenolysis is lactate (strenuous exercise)
Muscle and brain does not contain glu-6-phosphatase
Phosphorylase Pi
Glycogen Glu-1-P
Phosphoglucomutase
Glu –6-P
Glu-6-Phosphatase H2O
Pi
Glucose
DR S NAYAK 8
 Phosphorylase phosphorolytically splits -1,4
glucoside bonds from the outermost chains of
glycogen until 4 residues remain on either side of
– 1.6 branch point [limit dextrin]
  1.4 glucan transferase transfers 3 glucose
residue portion from one side chain to the other
exposing -1,6 branch points
 Amylo 1, 6 glucosidase splits the 1,6 linkages
 Acid maltase or -1,4-glucosidase (lysosomal enzyme)

degrades small quantity of glycogen. The significance of this
pathway is not clear
v Muscle glycogenolysis
Glycogen  Glu-1-P  Glu-6-P   glycolysis  lactate
DR S NAYAK 9
Glycogenolysis
DR S NAYAK 10
Regulation of glycogenesis and glycogenolysis
· The glycogen synthase and phosphorylase exist in

active and inactive forms
· The dephosphorylated form of glycogen synthase is
active
· Phosphorylated form of phosphorylase is active
The activation of phosphorylase depends on high
cAMP level. At the same time high cAMP level
inactivates glycogen synthase
Glycogen synthase b Phosphorylase a

H2O Protein phosphatase
Pi
Glycogen synthase a Phosphorylase b
(Glycogenesis ON) (Glycogenolysis OFF)
DR S NAYAK 11
Allosteric regulation
In a well fed state Glu–6– P level is high which
activates glycogen synthase
· On the other hand glu-6-p and ATP allosterically
inhibit phosphorylase
· Free glucose also act as inhibitor to phosphorylase
Glucose –6 –P ATP Liver glucose Muscle AMP

_ _ _
Glycogen Phosphorylase + Ca2+
Glycogen Glu-1-Phosphate
Glycogen Synthase
+
Glucose-6-phosphate
DR S NAYAK 12
Adrenaline (Liver and muscle)
Glucagon + (Liver only)
Adenylate Cyclase Adenylate Cyclase
ATP cAMP
PPi
Protein Kinase Protein Kinase
ATP ADP ATP ADP
Phosphorylase Phosphorylase Glycogen Glycogen
Kinase kinase Synthase (a) synthase (b)
Phosphorylase (b) Phosphorylase (a)
2ATP 2ADP
Glycogen Glucose-1-P
Pi
cAMP 51 AMP Glycogenesis ON
+ Phosphodiesterase
Insulin
DR S NAYAK 13
GLYCOGEN STORAGE DISEASES
Genetic diseases [may be inherited]
Deposition of abnormal type or increased quantity of glycogen in the tissues
Diseases Defect and Features
Type I. Glucose – 6 – phosphatase [liver]
Von Gierke’s disease Accumulation of glycogen in liver
Hypoglycaemia and ketosis
Type II. Lysosomal -1, 4 – glucosidase
Pompe’s disease Glycogen accumulates in
lysosomes, in all tissues
Enlarged liver and heart
Type III. Debranching enzyme [amylo -1,6-
Limit dextrinosis glucosidase
[Coris disease] Accumulation of polysaccharide
[limit dextrin] liver, heart, & muscle
TypeIV. Amylo pectinosis or Branching enzyme (glucosyl 4-6 transferase)
Andersons disease Accumulation of polysaccharide with few
branch points. Cirrhosis of liver
DR S NAYAK
14
Type V Muscle glycogen phosphorylase
McArdles disease Glycogen accumulates in the muscle
Diminished tolerance to exercise
Type VI. Liver glycogen Phosphorylase
Hers disease Liver enlarged
Von Gierke’s Disease
1. Fasting hypoglycemia
2. Lactic acidemia:Glucose is not synthesized from lactate produced in
muscle and liver. Lactate level increases and pH decreases
3. Hyperlipidemia: Block in gluconeogenesis leads to mobilisation fat to
meet energy requirement. So This increases free plasma FA & ketone
bodies.
4. Hyperuricemia: Accumulated glucose -6-p diverted to HMP pathway,
leading to increased synthesis of ribose and nucleotides, this enhances
metabolism of purine nucleotides and to uric acid later
5. Massive liver enlargement leads to cirrhosis
6. Children fail to grow
Given small quantity of food at frequent intervals
DR S NAYAK 15
REF: ESSENTIALS OF BIOCHEMISTRY: DR S NAYAK
16
THE
EXTRACELLULAR
MATRIX
Collagen
Dr. Neetu Mohan
St. Augustine
THE EXTRACELLULAR MATRIX
Collagen
Structural Elastin
Fibrillin
ECM Fibronectin,
Adhesive
PROTEINS laminin
Gel Forming Proteoglycans
Cell binding
Integrins
to ECM
RECALL: FIBROUS VS. GLOBULAR
PROTEINS
Globular proteins Fibrous proteins

• Spherical • Cylindrical
• Soluble in water • Low solubility in water
• Dynamic functions – • Shapes primarily due to
enzymes, receptors, secondary structure
antibodies • Structural functions –
collagen, elastin
RECALL: MOLECULES OF
IMPORTANCE
6 6
5 5
4 1 4 1
3 2 3 2
glucose galactose
RECALL: DISULPHIDE LINKAGE
FORMATION
H
+ -
H3 N C COO
CH2
SH
SH
CH2
+ -
H3 N C COO
H
RECALL: DISULPHIDE LINKAGE
FORMATION
H
+ -
H3 N C COO
CH2
S
Disulphide linkage
S
CH2
+ -
H3 N C COO
H
COLLAGEN
 approx. 25% of protein in
mammals
 Present in all tissues and
organs
 Fibrous
 Structural - extracellular
framework; strength
STRUCTURE OF COLLAGEN
 elongated
 3 α-chains (polypeptides) – approx. 1000 amino acids long
 each α-chain twisted into left-handed helix;
 each turn of the helix contains 3 amino acid residues
 three chains wound into right-handed superhelix
 rod-like; ~1.4nm diameter; ~300nm long
 triple helix; hydrogen bonds between chains
 hydroxyproline provides greater hydrogen bonding capability
PROCOLLAGEN
STRUCTURE OF COLLAGEN
 Amino acid sequence
 Pro and gly rich
 Pro causes bends in chain
 Gly every 3rd position
 Gly-X-Y
 X frequently pro (Approx. 100)
 Y frequently hydroxyproline (for rigidity) or
hydroxylysine (Approx. 100)
PROCOLLAGEN
Different types of Collagen; due to variation in amino acid sequence

e.g Type I contains two α-1 chains and one α-2 chain
Type II contains three α-1 chains
ORGANIZATION OF COLLAGENS
Type Structure Tissue Distribution Function
Fibril-forming Rope like Skin, bone, tendon, blood High tensile

Type I to III structure vessels, cornea (Type I) strength
Regularly Cartilage, vitreous body,
staggered intervertebral disks (Type
packing II)
Blood vessels, fetal skin
(Type III)
Network-Forming 3D mesh Basement membrane; Semi-

Types IV and VII Beneath stratified permeable
squamous epithelia filtration
barrier in
organs like
kidney and lung
Fibril-associated Bind to collagen Tendon, ligaments, Mechanical

Types IX and XII fibrils, linking them cartilage strength
BIOSYNTHESIS OF COLLAGEN (PART
1)
 Fibroblasts, osteoblasts etc.
 Nucleus : DNA transcribed into mRNA for α1 and/or α2; mRNA bound to
ribosomes in cytoplasm
 mRNA begins translation into polypeptide with signal sequence at the N terminal
 Signal seq directs polypeptide to rough ER
 Signal seq cleaved resulting in α1 and α2 chains
 Proline and lysine residues on polypeptide hydroxylated by prolyl and lysyl

hydroxylases respectively
 Hydroxylysine residues glycosylated
 Spontaneous formation of the triple helix flanked by non-helical N- and C-terminal
extensions of the pro-α chains (procollagen)
 Spontaneous formation due to interchain and intrachain disulphide bonds
between C terminal extensions
 Hydrogen bonds between hydroxyprolines
 Secretion of procollagen into the extracellular matrix
N-terminal C-terminal
(Lys and Pro)
(Hydroxylysine)
Hydrogen bonds between Disulphide Linkages

hydroxyprolines
BIOSYNTHESIS OF COLLAGEN (PART
2)
 Secretion of the procollagen molecule into extracellular matrix
 Cleavage by N and C-procollagen peptidases (remove terminal
extensions) – (tropocollagen)
 Cross-linking of tropocollagen molecules by lysyl oxidase
to form collagen fibrils
 Quarter staggered linking of collagen (Type I, II, III)
Cleavage of the N and C terminals by peptidases
Crosslinking by Lysyl Oxidase

Fibril-Forming Quarter-Staggered packing (Types 1 to 3)
NETWORK-FORMING COLLAGEN
(MESH)
 Mesh diagram
e.g Type IV collagen
POST-TRANSLATIONAL
MODIFICATION
 Hydroxylation of proline
Prolyl
H hydroxylase
H
COO- COO-
O2
HN C CO CH2 CH2 CO
HN C
H2C CH2 CH2 CH2
+ CO2 + H2C CH2
C Ascorbic
C
C ═O C00-
acid
H H
C00- H OH
Peptide-incorporated α-ketoglutarate succinate Peptide-incorporated

proline Note: Deficiency in ascorbic hydroxyproline
Acid (Vitamin C)
results in scurvy
POST-TRANSLATIONAL
MODIFICATION
 Hydroxylation of Lysine
H H
O O
═
═
N C C N C C
H Lysyl
CH2 H CH2
hydroxylase
COO- COO-
CH2 O2 CH2
CH2 CH2 +
CH2 + OH
CH2 CH2 H C
Ascorbic CO
2
acid
CH2 C ═O C00- CH2
+ C00- +
NH3 NH3
α-ketoglutarate succinate
Peptide-incorporated Note: Deficiency in ascorbic Peptide-incorporated
lysine Acid (Vitamin C) hydroxylysine
results in scurvy
POST-TRANSLATIONAL MODIFICATION
(CONT’D)
 Glycosylation of hydroxylysine
 Enzymatic
addition of
glucose or
glucosyl-galactose
 collagen galactosyltransferase
and collagen glucosyltransferase
CROSSLINKING
H H
O O
═
═
N C C N C C
H H CH2
CH2 Lysyl Oxidase
CH2 CH2
O2 NH3 + H2O
CH2 CH2
Oxidative deamination
CH2 C H
═
+ O
NH3
Lysine Allysine
Lysine
H H H
O O O
═
N C C N C C N C C
H (CH2)4 H H
(CH2)4 (CH2)4
Schiff
NH3+ Base N H2 N H
formation
═
C H H C H
O
═
C H (CH2)3 (CH2)3
H H
(CH2)3 N C C N C C
H ═
═
O O
N C C H H
═
O Schiff base
H
Allysine
Allysine
H
O
═
N C C
H
H O
(CH2)3
═
N C C
C H
H (CH2)2
═
O C C H
H2O
═
═
═
C H H C O
Aldol
(CH2)3 Condensation (CH2)3
H H
N C C N C C
═
═
O O
H H
Allysine
LYSYL OXIDASE AND COPPER
DEFICIENCY
 Lysyl oxidase
 Two identical subunits
 Encoded by LOX gene
 Contains a Cu2+ ion associated
with three histidine residues
Copper deficiency results in

impaired crosslinking
→ Abnormal collagen formation
ABNORMALITIES ASSOCIATED
WITH COLLAGEN
• Mutations in genes
• Abnormalities in enzymes
• Missing co-factors for enzymes
EHLERS-DANLOS SYNDROME (EDS)
 Inherited
 Autosomal dominant in mink, dogs, cats
 Autosomal recessive in cattle, sheep
 Dermatosparaxis
CAUSES:
 Deficiency of collagen
 Increased levels of collagenase (degrades collagen)
 2.5 times higher collagenase than in normal dogs
 Abnormal collagen
 Mutations in COL1A1, COL5A1 and COL5A2 genes (Types I and V collagen)
EHLERS-DANLOS SYNDROME (CONT’D)
Symptoms: Hyperextensibility of skin
• Stretchy skin
(hyperextensibility)
• Easy tearing of skin
• Very loose joints
(hypermobility)
• Cataracts
Joint hypermobility
CANINE OSTEOGENESIS IMPERFECTA
 Inherited (autosomal recessive)
 Canine inherited brittle bone disease
 Daschunds, golden retrievers, beagles
CAUSE:
 Abnormal collagen
 Mutation in COL1A2 gene
Symptoms:
 Brittle bones and teeth
 Loose joints
 Weak muscles and tendons
 Curved spine
CANINE CHONDRODYSPLASIA
 Canine dwarfism (osteochondrodysplasia)
 Inherited (autosomal dominant)
 Miniature poodles, cocker spaniels, Irish setters
CAUSE:
 mutation in integrin α subunit 10 gene
(ITGA10)
 integrin α subunit 10 is part of a receptor
which binds to collagen
Symptoms:
 Skeletal changes (short legs)
 Rounded end of bones
 Larger head
 Protruding lower jaw
 Crooked spine
JUNCTIONAL EPIDERMOLYSIS BULLOSA
 Inherited (autosomal recessive)
CAUSES:
 Abnormal collagen and laminin
 Mutations in COL17A1 gene
 Mutation JEB 1 (Belgian Draft horses)
 Mutation JEB 2 (American saddlebred horses)
Symptoms:
 Blisters in skin and mucosal membrane
 Blisters in gingiva and tongue
 Sloughing off of hooves
 Corneal lesions
 Dental dysplasia
SCURVY
CAUSE:
 Abnormality associated with ascorbic acid (Vit C) deficiency
 Guinea pigs and primates are unable to store Vit C
 Ascorbic acid- cofactor for prolyl and lysyl hydroxylase
 Hydroxylation of proline and lysine impaired
 Symptoms:
 Bleeding from mucous membranes
 Bruises on skin
 Spongy gums
THE
EXTRACELLULAR
MATRIX
Glycosaminoglycans
Dr. Neetu Mohan

St. Augustine
GLYCOSAMINOGLYCANS (GAGS)
 heteropolysaccharides
 Most abundant heteropolysaccharides in the body
 Long
 Unbranched
 Consist of repeating disaccharide units
 Negatively charged
 Bind large amounts of water to produce a gel-like matrix
 Found in mucus secretions
 Located primarily on the surface of cells or in the ECM
 Can be associated with protein to form proteoglycans
STRUCTURE OF GAGS
 The disaccharide units contain a modified
amino sugar and an acidic (uronic) sugar
 Amino sugar
 N-acetylgalactosamine (GalNAc)
 or N-acetylglucosamine (GlcNAc)
 Uronic acid
 glucuronate
 or iduronate
 highly negatively charged because of COO-

groups; some GAGs also contain SO2-4
groups
 extended in solution
 repel each other
 attract Na+ ions and therefore H2O
molecules
RECALL:
glucose
galactose
Negative
charges;
• Mucus
repulsion; slip secretions
past each
other • Synovial fluid
Extended in • Mucus secretions

solution • Synovial fluid
• Lubricating fluids
Low in joints
GAGs
compressibility
• Synovial fluids
• Vitreous humour
High viscosity
in joints
Rigidity
• Structural
integrity of cell
CLASSIFICATION OF GAGS
 GAGs may be classified according to:
1. Their monomeric composition
2. The type of glycosidic linkages
3. Degree and location of sulfate groups
 Six major types of GAGs:

 hyaluronic acid (hyaluronate)
 dermatan sulfate
 chondroitin sulfate
 heparin
 heparan sulfate
 keratan sulfate
DERMATAN SULFATE
6H 6CH OH
2
5 O 5 O
COO- -
O3SO H
H
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
L-iduronate N-acetyl-galactosamine-4-sulphate
CHONDROITIN 4 AND 6 SULFATES
6 6CH OH
COO- 2
5 O 5 O
D-glucuronate
N-acetyl-
H -
O3SO H
H galactosamine-
4 1 4 1
4-sulphate
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
6COO- 6CH OSO -

2 3
5 O 5 O N-acetyl-
D-glucuronate galactosamine-
H H
H OH 6-sulphate
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
KERATAN SULFATES I AND II
6CH OH 6CH OSO -
2 2 3
5 O 5 O N-acetyl-
D-galactose
H Glucosamine-
H
OH H 6-sulphate
4 1 4 1
O
H O H
H OH H
H
3 2 3 2
H OH H NHCOCH3
 D-galactose sugar instead of an acidic sugar
 Sulphate group may be present on C-6 of either
sugar
 most heterogenous GAG: may contain other
monosaccharides e.g
 L-fucose
 Mannose
 N-acetylneuraminic acid
HYALURONIC ACID
6COO- 6CH OH
2
5 O 5 O
H H
H H
4 1 4 1
O
OH H H H
H OH
3 2 3 2
O
H OH H NHCOCH3
D-glucuronate N-acetyl-glucosamine
 No sulphate groups
HEPARIN
6H 6CH OSO -
2 3
5 O 5 O
COO- H H H
H
4 1 4 1
O
O
OH H OH H
H
3 2 3 2
H OSO3- H NHOSO3-
L-iduronate-2-sulphate N-sulphoglucosamine-6-sulphate
 Glucosamine not acetylated; sulphate group
instead → sulfamide linkages
 Sulphates on C-3 or C-6 of glucosamine and
C-2 of uronic acid
HEPARAN SULFATES
 Glucosamine acetylated
 Fewer sulphate groups than heparin
GAG LOCATION SPECIAL PROPERTIES
Dermatan sulphate Skin, Blood vessels, Heart valves
Cornea (lie between collagen fibrils;
aids in corneal transparency)
Chondroitin-4-sulphate and Cartilage, Tendons, Aorta, Ligaments • Most abundant GAG in body
chondroitin-6-sulphate • Present in exoskeletons
• Forms proteoglycans with hyaluronic acid
and protein
• Bind collagen in cartilage, and hold fibers in a
tight, strong network
Keratan sulphate I and II KS II normally aggregated with chondroitin • Most heterogenous GAG: may contain other
sulphate in connective tissue, cartilage monosaccharides e.g
KS I found in cornea L-fucose
Mannose
N-acetylneuraminic acid
Heparin Arteries, Liver, Lungs, Skin • Anticoagulant
Heparan sulphate Extracellular, Basement membranes • Act as receptors

Cell surfaces • may participate in the mediation of cell growth
and cell communication
Hyaluronic acid synovial fluid of joints, vitreous humour of a component of non-covalently formed
eye, umbilical cord, loose connective tissue; complexes with proteoglycans in the ECM
cartilage their polysaccharides are very large
can displace a large volume of water
excellent lubricators and shock
absorbers
SYNTHESIS OF AMINO SUGARS – N-ACETYL-GLUCOSAMINE
O
═
-
6CH2OH 6CH2O P O
-
5 O
O 5 O
H H H hexokinase H Glucose-6-
H H
Glucose 4 4 4 phosphate
1 4 1
OH OH H OH
OH OH H OH
3 2 ATP ADP 3 2
H OH H OH
Glucose-6-phosphate
isomerase
O
═
- P O CH2OH
O OCH2
1
-
O
2
H H OH OH Fructose-6-
phosphate
4 3
OH H
D R . N . M O HA N
B S C , P H D B I O C HE M I S T R Y
O
Fructose-6-
═
phosphate
-
6CH2O P O
-
O O
5 O
═
- P O CH2OH H
O OCH2 H H
1 Fructose 6-phosphate
- 4 4 1 Glucosamine-
O transaminase
2 OH OH H OH6-
H H OH OH phosphate
3 2
4 3 H NH2 Acetyl Co A
OH H
O Coenzyme A
═
-
6CH2O P O Glucosamine
- phosphate
O
5 O N-acetyl
transferase
H H H
4 4 1
OH OH H OH
3 2 N-Acetyl
Glucosamine-
H NH 6-
C═ Ophosphate
CH3
Acetyl Co A
O
═
-
6CH2O P O 6CH2OH
-
O
5 O Phosphoacetylglucosamine 5 O
H mutase
H H H H H
4 4 1 4 4 1 O
═
-
OH OH H OH OH OH H O P O
3 N-Acetyl
2 3 2 - N-Acetyl
Glucosamine- O
H NH 6- H NH Glucosamine-
1-
C═ Ophosphate C═ O phosphate
CH3 CH3
UDP-N acetyl glucosamine
pyrophosphorylase O
6CH2OH UTP ║
C
HN
5 O UDP-N acetylglucosamine
H O═ C
H H
N
4 4 1 O O O O
═
═
═
═
OH OH H O P O -
O P O P O P OCH2
O
3 2 - - - -
O O O O
H NH
H H
C═ O
N-Acetyl
HO OH
CH3 Glucosamine-
1-
phosphate
(6C)
CMP- N-Acetyl-neuraminic acid UDP- N-Acetyl- Galactosamine

(9C) (6C)
SYNTHESIS OF AMINO SUGARS – N-ACETYL-GALACTOSAMINE
O
║
6CH2OH C
HN
5 O O═ C
H H H N UDP-N-
O O O acetylglucosamine
4 4 1
═
═
═
O
OH OH H O P O P O P OCH2
2 - - -
3 O O O
H NH H H
C═ O
HO OH
CH3
UDP-N-acetyl ║O
Glucosamine-4-
6CH2OH C
epimerase
HN
5 O O═ C
OH H H N
O O O UDP-N-
4 4 1
═
═
═
O acetylgalactosamine
H OH H O P O P O P OCH2
2 - - -
3 O O O
H NH H H
C═ O
HO OH
CH3
SYNTHESIS OF AMINO SUGARS – N-ACETYL-NEURAMINIC ACID
UDP-N-acetylglucosamine
UDP-N-acetyl
Glucosamine-2-
Epimerase/
N-acetylmannosamine
mannosamine ATP
kinase
ADP
N-acetylmannosamine-6-phosphate
N-acetyl
PEP + H2O
neuraminate-9-
Phosphate synthase Pi
N-Acetyl-neuraminic acid-9-phosphate
N-acetyl neuraminate-9-
phosphatase
N-Acetyl-neuraminic acid
CMP N-acetyl neuraminic
CTP
acid pyrophosphorylase
PPi
CMP- N-Acetyl-neuraminic acid
║
6CH2OH
HN
5 O
UDP-N- O═ C
H N
Acetyl H H
O O O
glucosamine 4 4 1
═
═
═
O
2 - - -
3 O O O
H NH H H
N-acetyl C═ O
mannosamine CH3
HO OH
O H
6CH2OH O ⑊ ̸
C1
═
-
5 O 6CH2O P O
H
- H3COCHN C H
H OH O 2
5 O
4 4 1 HO
NHCOCH3 H H OH
C H
OH OH
3
4 4 1
H
3 2 NHCOCH3 H C OH
ATP ADP OH OH 4
H H
2 H
3 H C OH
H H 5 O
═
-
N-acetylmannosamine CH2O P O
6
-6-phosphate O
-
-
O H phosphoenolpyruvate COO
⑊ ̸
C1 -
COO C═ O
O
H3COCHN C H
═
2 -
C O P O CH2
HO -
═
C H + O
3 CH2 H C OH
H C OH
4 + H3COCHN C H
H C OH HO
5 O H2O O C H
═
═
- -
CH2O P O O- P O
6 H C OH
- -
O O
H C OH
N-acetylmannosamine-6-phosphate O
═
-
CH2O P O
-
O
N-Acetyl-neuraminic
Acid-9-phosphate
COO- COO-
C═ O H
C═ O
H C OH
9
CH2 CH2
H C 8 OH
H C OH H C OH H C7 OH
H3COCHN C H H3COCHN C H H
6 O
H3C C N H COO-
HO C H HO 1
O C H
═
5 4 2
═
- - O
O P O H H
H C OH H C OH H OH
-
O 4 3
H C OH H C OH OH H
O
═
-
CH2O P O CH2OH
-
N-Acetyl-O
N-Acetyl- N-Acetyl-neuraminic acid
neuraminic
neuraminic
Acid
acid
-9-phosphate
6CH OH
2 SYNTHESIS OF ACIDIC SUGARS-
5 O
GLUCURONIC ACID
H OH
H Glucose-6-phosphate
4
Phosphoglucomutase
OH OH H
H
2 Glucose-1-phosphate
3 UDP
UDP-glucose
H OH PPi pyrophosphorylase
D-glucose
6COO- UDP-glucose
H2O 2NAD+ UDP-glucose
5 O dehydrogenase
NADH + H+
H OH
H UDP-glucuronate
4
OH OH H
H
2 Glucuronate
3
H OH
D-glucuronic acid
SYNTHESIS OF ACIDIC SUGARS-
IDURONIC ACID
 L-iduronic acid synthesis occurs after glucuronic acid is incorporated
into the polysaccharide
6COO-
6H
5 O Uronosyl-5- 5 O
H epimerase
H OH
H COO- OH
4
4
OH OH H
H OH OH H
H
3 2
3 2
H OH
H OH
D-glucuronic acid L-iduronic acid
PROTEOGLYCANS
 Also called
mucopolysaccharides
 GAGs linked to core proteins
 Core proteins linked to

hyaluronic acid (ionic
interactions; not covalent)
 Hyaluronic acid stabilized to

core proteins by link proteins
 GAGs extend perpendicularly

from the core in a “bottle
brush-like “structure
FUNCTIONS OF PROTEOGLYCANS
 Provide hydration
 Enables tissue to withstand compression
 e.g aggrecan in cartilage
PROTEOGLYCAN FUNCTION
Decorins • Regulates collagen fibril formation
• Modifying activity of transforming growth
factor beta
Perlecans • Charge selectivity in glomerular filtration
Syndicans and Glypicans • Binds to growth factors and ECM molecules

• Involved in signal transduction pathways that
regulate cell proliferation and cell shape
Heparan sulphate proteoglycans • Generation and differentiation of neurons;

synapse development
• Cell adhesion, motility, growth
RECALL: SERINE, THREONINE AND
ASPARAGINE
H H H
+ - + - + -
H3N C COO H3N C COO H3 N C COO
CH2 H3C C H CH2
OH OH C= O
Serine (Ser, S) Threonine (Thr, T) NH2
Asparagine(Asn, N)
RECALL: GALACTOSE AND XYLOSE
6CH OH
2 H
5 O 5 O
H H
OH OH H OH
4 1 4 1
H OH H OH OH H
H H
3 2 2
3
H OH H OH
GALACTOSE XYLOSE
PROTEOGLYCANS
 Linkage of GAGs to the core protein
 involves a specific trisaccharide
composed of two galactose and a xylose
residue attached to an amino acid from
the protein
 The trisaccharide is linked usually to a

Ser or Thr residue on the protein core
 This linkage is an O-glycosidic bond
 The protein cores of proteoglycans are

rich in Ser and Thr residues, which
allow multiple GAG attachments.
 Some forms of keratan sulfates are

linked to the protein core through an N-
glycosylamine bond to Asn
O-GLYCOSIDIC LINKAGE
xylosyltransferase
6H
5 C O
O Ser
H H
OH HO CH2 C
4 1
H N
OH OH H
H
3 2 Protein core
H OH
-D-xylose
H2O
O-linked N-acetylgalactosaminetransferase
6H
5 C O
O
Ser
OH H OH HO CH2 C
4 1
H N
H OH H
H
3 2 Protein core
H NH
C═ O
N acetylgalactosamine
CH3
H2O
N-GLYCOSYLAMINE LINKAGE
N-acetylglucosaminyltransferase
6H Asn
5 O H O C O
H H
OH N C CH2 C
4 1
H N
OH OH H H
H
3 2 Protein core
H NH
N-acetylglucosamine C═ O
CH3
H2O
SYNTHESIS OF PROTEOGLYCANS
 Protein core synthesized on rough ER
 Transported into rough ER
 UDP-xylose attached via a glycosidic
linkage to hydroxyl group of ser on
the protein core
 Enzyme which catalyzes this reaction
is xylosyltransferase
 Two galactose are then added to
xylose
 Glycosaminoglycan disaccharides are
then sequentially added
 Alternate amino and acidic sugar
 Some D-glucuronate converted to L-
iduronic acid
 Addition of sulphate groups
 Sulphate comes from 3’phosphoadenosyl-5’-phosphosulfate
 Sulfotransferase enzymes sulphate carbohydrate chains at specific sites
 Defects in sulfation cause several autosomal recessive disorders that
affect development and maintenance of the skeletal system
6 6CH OH
COO- 2
5 O 5 O
H -
O3SO H
H
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
DEGRADATION OF
GLYCOSAMINOGLYCANS
 Short half lives
 Hyaluronic acid – 3 days
 Chondroitin, dermatan sulfate – 10 days
 Keratan sulfate – 120 days*
 Phagocytosis of GAGs
 Engulfed by invagination of the cell membrane, forming a vesicle
 Vesicle fuses with a lysosome
 GAGs degraded in lysosomes

 Lysosomes contain hydrolytic enzymes that function at an optimal pH of 5 (acid
hydrolases)
 Lysosomal degradation of GAGs
 Firstly, the polysaccharide chains are cleaved by endoglycosidases, producing
oligosaccharides
 Further degradation of the oligosaccharides
LYSOSOMAL STORAGE DISEASES
 Impairments in lysosomal degradation → lysosomal storage diseases
(mucopolysaccharidoses)
 Defects in lysosomal enzymes responsible for GAG degradation

 accumulation of GAGs within cells
 There are at least 14 known types of lysosomal storage diseases that

affect GAG catabolism
 Common examples
 Hurler's syndrome
 Hunter's syndrome
 Sanfilippo syndrome
 Sly syndrome
 All of these disorders, except Hunter's syndrome, are inherited in an

autosomal recessive manner
Lysosomal Defective Enzyme Group/Sugar Affected Symptoms
Storage disease residue not GAG
removed or
affected
Hurler syndrome α-L-iduronidase L-iduronic acid Dermatan sulfate, Corneal clouding, mental
(cats and Plott heparan sulfate impairment, dwarfing, coarse
Hounds) facial features, upper airway
obstruction
Maroteux-Lamy N-acetylgalactosamine 4- Sulfate from N- Chondroitin sulfate, Corneal clouding, deafness,
Syndrome sulfatase acetylgalactosamine Dermatan sulfate pain due to compression of
(cats, miniature nerves
Pinscher dogs, rats)
Sly syndrome Β-glucuronidase Glucuronic acid Dermatan sulfate, Skeletal deformity,
(cats, dogs, mice) heparan sulfate, Short stature, corneal
Chondroitin4,6 clouding, mental deficiency
sulfate
San-Filippo syndrome Heparan sulfatase Sulfate from Heparan sulfate Severe nervous system
Type A glucosamine disorders;
(goats) Severe mental impairment
San-Filippo syndrome Acetyl co A Glucosaminide- Acetylglucosamine
Type C N-acyltransferase
(goats)
San-Filippo syndrome N-acetylglucosaminidase N-acetylglucosamine
Type B
(goats)
San-Filippo syndrome N-acetylglucosamine-6- Sulfate from N-
Type D sulfatase acetylglucosamine
(goats)
GLYCOPROTEINS
 Proteins with oligosaccharides attached
 (note: these oligosaccharides contain usually two to ten sugar residues)
 Linkage between the protein and oligosaccharide is covalent

 Either an O-glycosidic or N-glycosidic linkage
 The oligosaccharides chains:

 do not have disaccharide repeating units
 usually branched
 May or may not be negatively charged
 Contain highly variable amounts of carbohydrates

 E.g for immunoglobulin IgG, ˂ 4% of its mass is carbohydrate ; for human gastric
glycoproteins > 80% of its mass is carbohydrate
GLYCOPROTEINS
Function Glycoprotein
Structural Molecule Collagen
Lubricant and Protective Agent Mucins
Transport Molecule Transferrin, ceruloplasmin
Immunologic Molecule Immunoglobins, histocompatibility antigens
Enzyme Various, e.g alkaline phosphatase
Cell Attachment-recognition site Cell:cell (e.g sperm:oocyte; ZP3 glycoprotein in zona
pellucida); virus:cell (CXCR4, CCR5 to which HIV
binds); bacterium:cell
Cell adhesion Selectins
Receptor Various Proteins in hormone and drug action;
Chemokine receptors(CXCR4, CCR5 to which HIV
binds); dystrophin-glycoprotein (laminin receptor)
Hemostasis (and thrombosis) Prothrombin; Thrombin; Fibrinogen
ABO blood group antigens
Hormones HCG; TSH
GLYCOPROTEIN
SUGARS
Sugar Abbreviation
β-D-Glucose Glc
β-D-Galactose Gal
β-D-Mannose Man
α-L-Fucose Fuc
N-Acetylgalactosamine GalNAc
N-Acetylglucosamine GlcNAc
N-Acetylneuraminic acid (sialic acid) NeuNAc (Sia)
Xylose Xyl
GLYCOPROTEINS
 May be N-linked or O-linked
 N-linked oligosaccharides are attached via the amide nitrogens of
asparagine residues
 Complex glycoproteins
 High mannose glycoproteins
 O-linked oligosaccharides are attached to hydroxyl groups of serine,

threonine or hydroxylysine
 Membrane glycoproteins
 Extracellular glycoproteins
6H
5 C O
O
H H
OH HO CH2 C
4 1
H N
OH OH H
H
3 2
H OH
-D-xylose
H2O
O-LINKED GLYCOPROTEINS
N-GLYCOSIDIC LINKAGE
6H
5 O H O C O
H H
OH N C CH2 C
4 1
H N
OH OH H H
H
3 2
H NH
CH3
H2O
N-LINKED GLYCOPROTEINS
e.g lectins e.g ZP3 glycoprotein in zona pellucida

THE
EXTRACELLULAR
MATRIX
Glycosaminoglycans
Dr. Neetu Mohan

St. Augustine
GLYCOSAMINOGLYCANS (GAGS)
 heteropolysaccharides
 Most abundant heteropolysaccharides in the body
 Long
 Unbranched
 Consist of repeating disaccharide units
 Negatively charged
 Bind large amounts of water to produce a gel-like matrix
 Found in mucus secretions
 Located primarily on the surface of cells or in the ECM
 Can be associated with protein to form proteoglycans
STRUCTURE OF GAGS
 The disaccharide units contain a modified
amino sugar and an acidic (uronic) sugar
 Amino sugar
 N-acetylgalactosamine (GalNAc)
 or N-acetylglucosamine (GlcNAc)
 Uronic acid
 glucuronate
 or iduronate
 highly negatively charged because of COO-

groups; some GAGs also contain SO2-4
groups
 extended in solution
 repel each other
 attract Na+ ions and therefore H2O
molecules
RECALL:
glucose
galactose
Negative
charges;
• Mucus
repulsion; slip secretions
past each
other • Synovial fluid
Extended in • Mucus secretions

solution • Synovial fluid
Low in joints
GAGs
compressibility
• Synovial fluids
• Vitreous humour
High viscosity
in joints
Rigidity
• Structural
integrity of cell
CLASSIFICATION OF GAGS
 GAGs may be classified according to:
1. Their monomeric composition
2. The type of glycosidic linkages
3. Degree and location of sulfate groups
 Six major types of GAGs:

 hyaluronic acid (hyaluronate)
 dermatan sulfate
 chondroitin sulfate
 heparin
 heparan sulfate
 keratan sulfate
DERMATAN SULFATE
6H 6CH OH
2
5 O 5 O
COO- -
O3SO H
H
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
L-iduronate N-acetyl-galactosamine-4-sulphate
CHONDROITIN 4 AND 6 SULFATES
6 6CH OH
COO- 2
5 O 5 O
D-glucuronate
N-acetyl-
-
O3SO H
H H galactosamine-
4 1 4 1
4-sulphate
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
6COO- 6CH OSO -

2 3
5 O 5 O N-acetyl-
D-glucuronate galactosamine-
H H
H OH 6-sulphate
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
KERATAN SULFATES I AND II
6CH OH 6CH OSO -
2 2 3
5 O 5 O N-acetyl-
D-galactose Glucosamine-
H H
OH H 6-sulphate
4 1 4 1
O
H O H
H OH H
H
3 2 3 2
H OH H NHCOCH3
 D-galactose sugar instead of an acidic sugar
 Sulphate group may be present on C-6 of either
sugar
 most heterogenous GAG: may contain other
monosaccharides e.g
 L-fucose
 Mannose
 N-acetylneuraminic acid
HYALURONIC ACID
6COO- 6CH OH
2
5 O 5 O
H H
H H
4 1 4 1
O
OH H H H
H OH
3 2 3 2
O
H OH H NHCOCH3
D-glucuronate N-acetyl-glucosamine
 No sulphate groups
HEPARIN
6H 6CH OSO -
2 3
5 O 5 O
COO- H H
H H
4 1 4 1
O
O
OH H OH H
H
3 2 3 2
H OSO3- H NHOSO3-
L-iduronate-2-sulphate N-sulphoglucosamine-6-sulphate
 Glucosamine not acetylated; sulphate group
instead → sulfamide linkages
 Sulphates on C-3 or C-6 of glucosamine and
C-2 of uronic acid
HEPARAN SULFATES
 Glucosamine acetylated
 Fewer sulphate groups than heparin
GAG LOCATION SPECIAL PROPERTIES
Dermatan sulphate Skin, Blood vessels, Heart valves
Cornea (lie between collagen fibrils;
aids in corneal transparency)
Chondroitin-4-sulphate and Cartilage, Tendons, Aorta, Ligaments • Most abundant GAG in body
chondroitin-6-sulphate • Present in exoskeletons
• Forms proteoglycans with hyaluronic acid
and protein
• Bind collagen in cartilage, and hold fibers in a
tight, strong network
Keratan sulphate I and II KS II normally aggregated with chondroitin • Most heterogenous GAG: may contain other
sulphate in connective tissue, cartilage monosaccharides e.g
KS I found in cornea L-fucose
Mannose
N-acetylneuraminic acid
Heparin Arteries, Liver, Lungs, Skin • Anticoagulant
Heparan sulphate Extracellular, Basement membranes • Act as receptors

Cell surfaces • may participate in the mediation of cell growth
and cell communication
Hyaluronic acid synovial fluid of joints, vitreous humour of a component of non-covalently formed
eye, umbilical cord, loose connective tissue; complexes with proteoglycans in the ECM
cartilage their polysaccharides are very large
can displace a large volume of water
excellent lubricators and shock
absorbers
SYNTHESIS OF AMINO SUGARS – N-ACETYL-GLUCOSAMINE
O
═
-
6CH2OH 6CH2O P O
-
5 O
O 5 O
H H H hexokinase H Glucose-6-
H H
Glucose 4 4 4
phosphate
1 4 1
OH OH H OH
OH OH H OH
3 2 ATP ADP 3 2
H OH H OH
Glucose-6-phosphate
isomerase
O
═
- P O CH2OH
O OCH2
1
-
O
2
H H OH OH Fructose-6-
phosphate
4 3
OH H
DR. N. MOHAN
B S C , P H D B I O C H E M I S TR Y
O
Fructose-6-
═
phosphate
-
6CH2O P O
-
O O
5 O
═
- P O CH2OH H
O OCH2 H H
1 Fructose 6-phosphate
- 4 4 1 Glucosamine-
O transaminase
2 OH OH H OH6-
H H OH OH phosphate
3 2
4 3 H NH2 Acetyl Co A
OH H
O Coenzyme A
═
-
6CH2O P O Glucosamine
- phosphate
O
5 O N-acetyl
transferase
H H H
4 4 1
OH OH H OH
3 2 N-Acetyl
Glucosamine-
H NH 6-
C═ Ophosphate
CH3
Acetyl Co A
O
═
-
6CH2O P O 6CH2OH
-
O
5 O Phosphoacetylglucosamine 5 O
H mutase
H H H H H
4 4 1 4 4 1 O
═
-
OH OH H OH OH OH H O P O
3 N-Acetyl
2 3 2 - N-Acetyl
Glucosamine- O
H NH 6- H NH Glucosamine-
1-
C═ Ophosphate C═ O phosphate
CH3 CH3
UDP-N acetyl glucosamine
pyrophosphorylase O
6CH2OH UTP ║
C
HN
5 O UDP-N acetylglucosamine
H O═ C
H H
N
4 4 1 O O O O
═
═
═
═
OH OH H O P O -
O P O P O P OCH2
O
3 2 - - - -
O O O O
H NH
H H
C═ O
N-Acetyl
HO OH
CH3 Glucosamine-
1-
phosphate
(6C)
CMP- N-Acetyl-neuraminic acid UDP- N-Acetyl- Galactosamine

(9C) (6C)
SYNTHESIS OF AMINO SUGARS – N-ACETYL-GALACTOSAMINE
O
║
6CH2OH C
HN
5 O O═ C
H H H N UDP-N-
4 4 1
O O O acetylglucosamine
═
═
═
O
2 - - -
3 O O O
H NH H H
C═ O
HO OH
CH3 UDP-N-acetyl ║O
Glucosamine-4-
6CH2OH C
epimerase
HN
5 O O═ C
OH H H N
4 4 O O O UDP-N-
1
═
═
═
O acetylgalactosamine
H OH H O P O P O P OCH2
2 - - -
3 O O O
H NH H H
C═ O
HO OH
CH3
SYNTHESIS OF AMINO SUGARS – N-ACETYL-NEURAMINIC ACID
UDP-N-acetylglucosamine
UDP-N-acetyl
Glucosamine-2-
Epimerase/
N-acetylmannosamine
mannosamine ATP
kinase
ADP
N-acetylmannosamine-6-phosphate
N-acetyl
PEP + H2O
neuraminate-9-
Phosphate synthase Pi
N-Acetyl-neuraminic acid-9-phosphate
N-acetyl neuraminate-9-
phosphatase
N-Acetyl-neuraminic acid
CMP N-acetyl neuraminic
CTP
acid pyrophosphorylase
PPi
CMP- N-Acetyl-neuraminic acid
║
6CH2OH
HN
5 O
UDP-N- O═ C
Acetyl H H H N
O O O
glucosamine 4 4 1
═
═
═
O
2 - - -
3 O O O
H NH H H
N-acetyl C═ O
mannosamine CH3
HO OH
O H
6CH2OH O ⑊ ̸
C1
═
-
5 O 6CH2O P O
H
- H3COCHN C H
H OH O 2
5 O
4 4 1 HO
NHCOCH3 H H OH
C H
OH OH
3
4 4 1
H
3 2 NHCOCH3 H C OH
ATP ADP OH OH 4
H H
2 H
3 H C OH
H H 5 O
═
-
N-acetylmannosamine CH2O
6
P O
-6-phosphate O
-
-
O H phosphoenolpyruvate COO
⑊ ̸
C1 -
COO C═ O
O
H3COCHN C H
═
-
2 C O P O CH2
HO
═
-
C H + O
3 CH2 H C OH
H C OH
4 + H3COCHN C H
H C OH HO
5 O H2O O C H
═
═
- -
CH2O P O O- P O
6 H C OH
- -
O O
H C OH
N-acetylmannosamine-6-phosphate O
═
-
CH2O P O
-
O
N-Acetyl-neuraminic
Acid-9-phosphate
-
COO COO
-
C═ O C═ O
H
H C OH
9
CH2 CH2
H C 8 OH
H C OH H C OH H C7 OH
H3COCHN C H H3COCHN C H H
6 O
H3 C C N H COO-
HO C H HO 1
═
O C H 5 4
═
2
- - O
O P O H H
H C OH H C OH H OH
-
O 4 3
H C OH H C OH OH H
O
═
-
CH2O P O CH2OH
-
N-Acetyl-O
N-Acetyl- N-Acetyl-neuraminic acid
neuraminic
neuraminic
Acid
acid
-9-phosphate
6CH OH
2 SYNTHESIS OF ACIDIC SUGARS-
5 O
GLUCURONIC ACID
H OH
H Glucose-6-phosphate
4
Phosphoglucomutase
OH OH H
H
2 Glucose-1-phosphate
3 UDP
UDP-glucose
H OH PPi pyrophosphorylase
D-glucose
6COO- UDP-glucose
H2O 2NAD+ UDP-glucose
5 O dehydrogenase
NADH + H+
H OH
H UDP-glucuronate
4
OH OH H
H
2 Glucuronate
3
H OH
D-glucuronic acid
SYNTHESIS OF ACIDIC SUGARS-
IDURONIC ACID
 L-iduronic acid synthesis occurs after glucuronic acid is incorporated
into the polysaccharide
6COO-
6H
5 O Uronosyl-5- 5 O
H epimerase
H OH
H COO- OH
4
4
OH OH H
H OH OH H
H
3 2
3 2
H OH
H OH
D-glucuronic acid L-iduronic acid
PROTEOGLYCANS
 Also called
mucopolysaccharides
 GAGs linked to core proteins
 Core proteins linked to

hyaluronic acid (ionic
interactions; not covalent)
 Hyaluronic acid stabilized to

core proteins by link proteins
 GAGs extend perpendicularly

from the core in a “bottle
brush-like “structure
 Provide hydration
 Enables tissue to withstand compression
 e.g aggrecan in cartilage
PROTEOGLYCAN FUNCTION
Decorins • Regulates collagen fibril formation
• Modifying activity of transforming growth
factor beta
Perlecans • Charge selectivity in glomerular filtration
Syndicans and Glypicans • Binds to growth factors and ECM molecules

• Involved in signal transduction pathways that
regulate cell proliferation and cell shape
Heparan sulphate proteoglycans • Generation and differentiation of neurons;

synapse development
• Cell adhesion, motility, growth
RECALL: SERINE, THREONINE AND
ASPARAGINE
H H H
+ - + - + -
H3N C COO H3N C COO H3 N C COO
CH2 H3C C H CH2
OH OH C= O
Serine (Ser, S) Threonine (Thr, T) NH2
Asparagine(Asn, N)
RECALL: GALACTOSE AND XYLOSE
6CH OH
2 H
5 O 5 O
H H
OH OH H OH
4 1 4 1
OH H
H H OH OH H
H
3 2 2
3
H OH H OH
GALACTOSE XYLOSE
PROTEOGLYCANS
 Linkage of GAGs to the core protein
 involves a specific trisaccharide
composed of two galactose and a xylose
residue attached to an amino acid from
the protein
 The trisaccharide is linked usually to a

Ser or Thr residue on the protein core
 This linkage is an O-glycosidic bond
 The protein cores of proteoglycans are

rich in Ser and Thr residues, which
allow multiple GAG attachments.
 Some forms of keratan sulfates are

linked to the protein core through an N-
glycosylamine bond to Asn
xylosyltransferase
6H
5 C O
O Ser
H H
OH HO CH2 C
4 1
H N
OH OH H
H
3 2 Protein core
H OH
-D-xylose
H2O
O-linked N-acetylgalactosaminetransferase
6H
5 C O
O
Ser
OH H OH HO CH2 C
4 1
H N
H OH H
H
3 2 Protein core
H NH
C═ O
N acetylgalactosamine
CH3
H2O
N-GLYCOSYLAMINE LINKAGE
N-acetylglucosaminyltransferase
6H Asn
5 O H O C O
H
H OH N C CH2 C
4 1
H N
OH OH H H
H
3 2 Protein core
H NH
CH3
H2O
 Protein core synthesized on rough ER
 Transported into rough ER
 UDP-xylose attached via a glycosidic
linkage to hydroxyl group of ser on
the protein core
 Enzyme which catalyzes this reaction
is xylosyltransferase
 Two galactose are then added to
xylose
 Glycosaminoglycan disaccharides are
then sequentially added
 Alternate amino and acidic sugar
 Some D-glucuronate converted to L-
iduronic acid
 Addition of sulphate groups
 Sulphate comes from 3’phosphoadenosyl-5’-phosphosulfate
 Sulfotransferase enzymes sulphate carbohydrate chains at specific sites
 Defects in sulfation cause several autosomal recessive disorders that
affect development and maintenance of the skeletal system
6 6CH OH
COO- 2
5 O 5 O
H -
O3SO H
H
4 1 4 1
O
OH H H H
H H
3 2 3 2
O
H OH H NHCOCH3
DEGRADATION OF
GLYCOSAMINOGLYCANS
 Short half lives
 Hyaluronic acid – 3 days
 Chondroitin, dermatan sulfate – 10 days
 Keratan sulfate – 120 days*
 Phagocytosis of GAGs
 Engulfed by invagination of the cell membrane, forming a vesicle
 Vesicle fuses with a lysosome
 GAGs degraded in lysosomes

 Lysosomes contain hydrolytic enzymes that function at an optimal pH of 5 (acid
hydrolases)
 Lysosomal degradation of GAGs
 Firstly, the polysaccharide chains are cleaved by endoglycosidases, producing
oligosaccharides
 Further degradation of the oligosaccharides
LYSOSOMAL STORAGE DISEASES
 Impairments in lysosomal degradation → lysosomal storage diseases
(mucopolysaccharidoses)
 Defects in lysosomal enzymes responsible for GAG degradation

 accumulation of GAGs within cells
 There are at least 14 known types of lysosomal storage diseases that

affect GAG catabolism
 Common examples
 Hurler's syndrome
 Hunter's syndrome
 Sanfilippo syndrome
 Sly syndrome
 All of these disorders, except Hunter's syndrome, are inherited in an

autosomal recessive manner
Lysosomal Defective Enzyme Group/Sugar Affected Symptoms
Storage disease residue not GAG
removed or
affected
Hurler syndrome α-L-iduronidase L-iduronic acid Dermatan sulfate, Corneal clouding, mental
(cats and Plott heparan sulfate impairment, dwarfing, coarse
Hounds) facial features, upper airway
obstruction
Maroteux-Lamy N-acetylgalactosamine 4- Sulfate from N- Chondroitin sulfate, Corneal clouding, deafness,
Syndrome sulfatase acetylgalactosamine Dermatan sulfate pain due to compression of
(cats, miniature nerves
Pinscher dogs, rats)
Sly syndrome Β-glucuronidase Glucuronic acid Dermatan sulfate, Skeletal deformity,
(cats, dogs, mice) heparan sulfate, Short stature, corneal
Chondroitin4,6 clouding, mental deficiency
sulfate
San-Filippo syndrome Heparan sulfatase Sulfate from Heparan sulfate Severe nervous system
Type A glucosamine disorders;
(goats) Severe mental impairment
San-Filippo syndrome Acetyl co A Glucosaminide- Acetylglucosamine
Type C N-acyltransferase
(goats)
San-Filippo syndrome N-acetylglucosaminidase N-acetylglucosamine
Type B
(goats)
San-Filippo syndrome N-acetylglucosamine-6- Sulfate from N-
Type D sulfatase acetylglucosamine
(goats)
GLYCOPROTEINS
 Proteins with oligosaccharides attached
 (note: these oligosaccharides contain usually two to ten sugar residues)
 Linkage between the protein and oligosaccharide is covalent

 Either an O-glycosidic or N-glycosidic linkage
 The oligosaccharides chains:

 do not have disaccharide repeating units
 usually branched
 May or may not be negatively charged
 Contain highly variable amounts of carbohydrates

 E.g for immunoglobulin IgG, ˂ 4% of its mass is carbohydrate ; for human gastric
glycoproteins > 80% of its mass is carbohydrate
GLYCOPROTEINS
Function Glycoprotein
Structural Molecule Collagen
Lubricant and Protective Agent Mucins
Transport Molecule Transferrin, ceruloplasmin
Immunologic Molecule Immunoglobins, histocompatibility antigens
Enzyme Various, e.g alkaline phosphatase
Cell Attachment-recognition site Cell:cell (e.g sperm:oocyte; ZP3 glycoprotein in zona
pellucida); virus:cell (CXCR4, CCR5 to which HIV
binds); bacterium:cell
Cell adhesion Selectins
Receptor Various Proteins in hormone and drug action;
Chemokine receptors(CXCR4, CCR5 to which HIV
binds); dystrophin-glycoprotein (laminin receptor)
Hemostasis (and thrombosis) Prothrombin; Thrombin; Fibrinogen
ABO blood group antigens
Hormones HCG; TSH
GLYCOPROTEIN
SUGARS
Sugar Abbreviation
β-D-Glucose Glc
β-D-Galactose Gal
β-D-Mannose Man
α-L-Fucose Fuc
N-Acetylgalactosamine GalNAc
N-Acetylglucosamine GlcNAc
N-Acetylneuraminic acid (sialic acid) NeuNAc (Sia)
Xylose Xyl
GLYCOPROTEINS
 May be N-linked or O-linked
 N-linked oligosaccharides are attached via the amide nitrogens of
asparagine residues
 Complex glycoproteins
 High mannose glycoproteins
 O-linked oligosaccharides are attached to hydroxyl groups of serine,

threonine or hydroxylysine
 Membrane glycoproteins
 Extracellular glycoproteins
6H
5 C O
O
H H
OH HO CH2 C
4 1
H N
OH OH H
H
3 2
H OH
-D-xylose
H2O
O-LINKED GLYCOPROTEINS
N-GLYCOSIDIC LINKAGE
6H
5 O H O C O
H
H OH N C CH2 C
4 1
H N
OH OH H H
H
3 2
H NH
CH3
H2O
e.g lectins e.g ZP3 glycoprotein in zona pellucida

Metalloenzymes
Dr. Neetu Mohan

School of Veterinary Medicine,
Faculty of Medical Sciences,
University of the West Indies, St. Augustine
Metal ions in catalysis
• One third of all enzymes require a metal ion for catalytic function
Na+, K+ Mg2+,Ca2+ Zn2+, Ni2+ Fe, Cu, Co, Mo, Mn
Favored +1 +2 +2 Variable, more

Oxidation than one state
state
Stability of Very low Low to High High (medium

complex medium for Mn2+ and
Fe2+)
After Frausto de Silva and Williams

Metal Co-factors of Enzymes
Metal Co-factor Enzyme Class

Magnesium Kinases
Calcium Hydrolases
Potassium Pyruvate kinase
Iron Cytochromes
Zinc DNA binding
Copper Oxidases
Manganese Oxidases
Cobalt Mutases
Selenium Peroxidases
Metalloenzymes vs. Metal Activated Enzymes
Metalloenzyme Metal - activated

Metal permanently bound to Enzyme Metal not permanently bound
Metal stays bound, unless removed by Metal easily removed during enzyme
chelator isolation
Fixed number of metals per enzyme Number not fixed
Coordinate covalent bonding Electrostatic bonding
Limited number of binding sites (usually Multiple binding sites
one)
Mostly transition metals (Zn2+, Fe2+, Mostly group 1 and 2 metals (Na+, K+,
Cu2+, Co2+) Mg2+, Ca2+)
Coordinate covalent bonding in
hemoglobin
• Fe has 6 coordination
positions
• 4 of 6 of Fe's coordination
positions are occupied by
bonds to the porphyrin ring
• a fifth coordination position is
bonded to a histidine side
chain of the protein
• the sixth is bonded with
oxygen
Zn
 Approx. 300 enzymes require zinc
 Transcription and Translation

 DNA, RNA polymerases
 Buffer system
 Carbonic anhydrase
 Others
 Carboxypeptidase
 Aminopeptidase
 Collagenase
Fe
• Electron Transport & Energy Metabolism

 Cytochromes – electron carriers in membranes e.g cytochrome
oxidase
 Fe-S proteins – electron carriers
• Others
 Catalase- enzyme that breaks down H2O2 (hydrogen peroxide)
 Ribonucleotide reductase - forms deoxyribose from ribose
Electron Transport Complexes
 ETC Enzymes contain one or more of the following:
 Heme, which contains Fe. (Proteins containing heme are
normally called cytochromes). Fe switches between oxidation
states 3+ and 2+
 Iron-Sulfur clusters (Fe-S)
 Copper. Cu switches between oxidation states 2+ and 1+
heme
Electron Transport Complexes
..
.. Cyt
.. .. Cyt b Cyt c1 Cyt c a+a3
NADH FMN CoQH2 (Fe3+) (Fe2+) (Fe3+) (Fe2+) O2
NAD+ FMNH2 CoQ Cyt b Cyt c1 Cyt c Cyt H2 O

.. (Fe2+) (Fe3+) (Fe2+) a+a3 ..
.. .. (Fe3+)
Cu
 Cytochrome oxidase
 Electron transfer; O2 final electron acceptor
 Lysyl oxidase
 Crosslinking of collagen
 Copper-Zinc superoxide dismutase
 Free radical scavenger enzyme
Metal-activated enzymes
 Eg. Phosphotransferases
 ATP→ ADP + Pi
 Mg binds to ATP to form Mg-
ATP
 Mg-ATP binds to the
phosphotransferase enzyme
to form a Mg –ATP- enzyme
complex
 Electrostatic interactions

Biochem Combined

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Objectives

The student should be able to:

 Describe aerobic and anaerobic glycolysis

Glycolysis: Degradation of glucose to pyruvate (Lactate under

• Carbohydrates are the important energy source of the

Importance of the pathway

• Most of the reactions of glycolysis are reversible

• The entry of glucose from ECF to cell (ICF) is under

• Glycolysis occurrence is the pre-requisite for the

• Aerobic oxidation takes place in the cells possessing

• Glycolysis is the major pathway for ATP synthesis in

•Glucose 6-P: Impermeable to the cell membrane.

3. PFK is an allosteric inducible key enzyme and the reaction is

5. Reversible,end product contains a high-energy

6. The high energy of 1,3 bisphosphoglycerate is

8. Reversible reaction, Mg2+ act as activator

9. High energy of Phosphoenolpyruvate is trapped into

10. If anaerobic conditions prevail, the reoxidation of NADH

• Glycolysis is the major source of energy in anaerobiosis

Pyruvate kinase is an inducible enzyme that increases its synthesis

F 2,6 BP is the regulatory factor for controlling PFK and then

The function of synthesis and degradation of Fructose 2, 6-BP is brought out by

The activity of these enzymes is controlled by covalent modification. cAMP

There will be no stimulation when Fru-2, 6 bisphosphate decreases, with

 No energy is trapped from 2, 3 BPG

 The BPG when combines with Hb, reduces the affinity

 Therefore the 2, 3 BPG increases in hypoxic condition

 In hexokinase deficiency phosphorylation does not

Lactic acidosis is caused by:

At the end of this lecture student should be able to :

Describe the integration of metabolism during fed and fasting conditions

DR. SHIVANANDA NAYAK 1

DR. SHIVANANDA NAYAK

Fatty acid oxidation:

Amino acid degradation:

Hexose monophosphate shunt:

DR. SHIVANANDA NAYAK

Glycogen metabolism: Glycogen is the storage form of glucose,

1.The availability of substrates

DR. SHIVANANDA NAYAK

The various tissues and organs of the body work in a well-coordinated

It is a metabolic stress, which imposes certain

DR. SHIVANANDA NAYAK 12

Glucose is the fuel of choice for brain and muscle.

 Protein meet the fuel demands of the body

Starvation associated with decreased insulin and

•FA and TG synthesis completely stopped here.

In the early phase of starvation, the brain mostly

The Ketones bodies play a central role in prolonged

Ref: Essentials of Biochemistry

Dr. Shivananda Nayak

Professor Shivananda Nayak

• Conditions in which ketone bodies are formed

Acetoacetate, -hydroxy butyrate and acetone are collectively called

In starvation TCA cycle is impaired due to the deficiency of

Therefore acetyl CoA converted to ketone bodies to meet the

Because of the lack of insulin the carbohydrate metabolism is impaired

Feeding high fat diet:

Excess breakdown of fatty acids in the liver takes place.

β-OH β-methyl glutaryl CoA [HMG CoA]

 -hydroxy butyrate CO2+ Acetone

Utilisation of ketone bodies (ketolysis)

Acetoacetate and -hydroxybutyrate can be used as a source of energy

During prolonged starvation brain utilize ketone bodies.