VNUHCM SCHOOL OF MEDICINE

ENZYMES- VITAMINS-
HORMONES

Lecturer: Huynh Thi Dieu Hien

Email: htdhien@medvnu.edu.vn
PN: 090 68 444 58
CHAPTER 3: THE PHYSIOLOGIC ROLES OF
ENZYMES
REFERENCES:
1/ Turco, S. & Reichenbecher, V. USMLE Step 1,
Biochemistry and Medical Genetics (lecture notes).
Kaplan Medical. 2013.
2/ Richard G. Macdonald William G. Chaney, Road Map
Biochemistry, University of Nebraska medical center Omaha.
3/ John W bayer, Marek H. Dominiczak , Medical
Biochemistry, Elsevier, 2019.
What is an Enzyme?
Role of Enzymes?
Enzyme activation?
 Learning objectives
- 1/ Describe the characteristics of enzymatic reactions from the viewpoint of

energy, equilibrium & kinetics.

- 2/ Discuss the structure and composition of enzymes, including the role of

cofactors, and conditions that affect enzymatic reactions.

- 3/ Describe enzyme kinetics.

- 4/ Describe the elements of enzyme structure substrate specificity & catalytic

activity.

- 5/ Describe regulatory mechanisms affecting enzymatic reactions.

6/ Discuss the therapeutic use of enzyme inhibitors and the diagnostic utility

of clinical enzyme assays.

- Approximately 1300 different enzymes found in the
human cell, help us to see, hear, feel, move, digest
food, and think.

ORDER

EFFICIENT
METABOLISM

BRANCHING
METABOLIC SEQUENCE
PATHWAYS
- Physiologic reactions: absence of enzymes in the
body > life would be impossible.
- Almost all enzymes are proteins, although some
RNAs (ribozymes).
- In clinic: measure substrate concentrations in
biological fluids (serum, plasma), and
measurement of enzyme concentrations helpful in
the diagnosis and monitoring of many clinical
conditions.
I. ENZYMATIC REACTION:
I.1. Factors affecting enzymatic reactions
 - Cytosolic
enzymes: 7-8.
- Pepsin: 1.5–2.0
- Trypsin and
chymotrypsin
have alkaline
pH optima.
- Lysosomal
enzymes: acidic
PH.
I. 2. Enzyme activity
- Enzyme unit:
+ IU or U, also E, Units, International unit of enzymes is
commonly used as the standard unit of activity (vitamin, vaccine,
hormone,...).
One international unit (IU) of enzyme catalyzes conversion
of 1 µmol of substrates to product per minute.
1 IU= 1μmol/min.
+ Katal units: 1 mole of substrates into 1 mole of product per
second (1 kat = 1 mol/s).
Examples:
+ Insulin: 1 IU = 0,0347 mg insulin of human = 45,5 μg pure
insulin.
+ Vitamin A: 1IU = 0,3 μg retinol = 0,6 μg beta- carotene.
+ Vitamin C: 1IU= 50 μg L-ascorbic acid.
Convert Enzyme unit to Katal (U to kat):
https://www.convert-measurement-units.com
- The specific activity of an enzyme is a measure of the
number of IU/mg protein (solid) or IU/ml (liquid).
µmol/min/mg of protein or IU/mg of protein
- The specific activity of enzymes varies greatly among
tissues. The enzymes for cholesterol synthesis, for
example, have a higher specific activity (IU/mg tissue)
in liver than in muscle, consistent with the role of the
liver in the biosynthesis of cholesterol.
I.3 Reaction and substrate specificity
- Most enzymes are highly specific: catalyzation and
substrates.
+ catalytic site: amino acid residues.
+ Substrate specificity: size, structure, charges, polarity,
and hydrophobicity of the substrate-binding site.
Reaction profile for enzymatic
and nonenzymatic reactions.

The transition state of the

enzyme-catalyzed reaction has a
lower energy than that of the
uncatalyzed reaction, the reaction
can proceed faster. ES complex,
enzyme–substrate complex; EP
complex, enzyme–
productcomplex.
I.4 Enzyme classification
- Highly specific enzymes  one specific chemical
reaction: ex. catalase and urease, which degrade H2O2
and urea.

- Some enzymes have broader substrate specificity. Ex,

the pancreatic enzymes chymotrypsin, trypsin, and
elastase, that contains a reactive serine residue in the
catalytic site. They catalyze the hydrolysis of peptide
bonds on the carboxyl side of a limited range of amino
acids in protein. They have similar structures and
catalytic mechanisms, their substrate specificities are
quite different because of structural features of the
substrate binding site (Fig. 2).
 Fig 2. Characteristics of the substrate binding sites in
the serine proteases chymotrypsin, trypsin, and
elastase.
In chymotrypsin, a hydrophobic pocket binds aromatic amino acid residues such as
phenylalanine (Phe). In trypsin, the negative charge of the aspartate residue in the
substrate-binding site promotes cleavage to the carboxyl side of positively charged
lysine (Lys) and arginine (Arg) residues. In elastase, side chains of valine and threonine
block the substrate-binding site and permit binding of amino acids with small or no
side chains, such as glycine (Gly). ▼, site of hydrolysis by enzyme.
Serine hydrolases: largest known enzyme classes, ~200 enzymes or

1% of the genes in the human proteome.

- Presence of a particular serine at the active site.

- The hydrolysis of the ester or peptide bond proceeds in two steps.

+ First, the acyl (R1-C=O) part of the substrate (the acid part of an ester

or the part of a peptide ending in a carboxyl group) is transferred to the

serine, making a new ester or amide bond and releasing the other part of

the substrate (the alcohol of an ester or the part of the peptide ending in

an amino group) is released.

+ Later, in a slower step, the bond between the serine and the acyl group

is hydrolyzed by water or hydroxide ion, regenerating free enzyme.

The activities of some tissue-specific enzymes and
isozymes (are enzymes that differ in amino acid sequence
but catalyze the same chemical reaction) are measured in
serum for diagnostic purposes.
 Isozymes

EX: LDH (Lactate

Dehydrogenase)
Ex: the enzyme LDH

The 5 isozymes of LDH.

 Densitometric patterns of the lactate dehydrogenase (LDH)
isozymes in the serum of patients diagnosed with myocardial
infarction or acute hepatitis
Self- study

Isozyme: glucokinase, creatine phosphokinase,

alkaline phosphatase.
II. Structure of enzyme

- Apoenzyme
- Holoenzyme
- Coenzyme
- Cofactor
- Catalytic site

Activation of the enzyme

Cofactor Examples:

- Mg2+: Hexokinase, Enolase, Glucose 6-P.

- Zn2+: Alcohol dehydrogenase, Carboxypeptidase etc.

- Pyridoxine (B6): Pyridoxal phosphate is a coenzyme of pyridoxine

precursor of vitamin B6 that carries out amino group transfer
- Activator metal ions: loosely bound.
- Metal ions of metalloenzymes: tightly bound.
- Coenzymes: loosely bound.
- Prosthetic groups: tightly bound.
Metal ions such as Cu2+, Fe3+, Mg2+, Mn2+, Mo2+,
Ni2+, Zn 2+,...

Organic Cofactor
Coenzymes

- Coenzyme or Helper molecules are small organic

molecules that are required for activity of certain enzymes.
-They bind loosely to an enzyme at the active site to help
catalyze reactions.
-Most coenzymes are vitamin derivatives.
 III.. Kinetics of Enzyme-Catalyzed Reactions
- The rate or velocity : change in concentration of reactants or products
with time.
1. Velocity decreases as reactants are used up to the point of
equilibrium rate = 0.
2. The rates of most physiologic reactions depend only on the
concentration of one reactant.
a. Such reactions are said to obey first-order kinetics.
b. Progress of such reactions can be followed according to the half-life
of that reactant.
- The energy difference during conversion of reactants to products in a
reaction can be represented by an energy diagram (Fig. Bellow).
 1. The activation energy is the energy barrier that must be overcome to
convert the reactants to products.

2. As the reaction progresses and if sufficient activation energy is

available, a state of high energy termed the transition state is reached; this
state has a structure intermediate between reactants and products.
3. For a chemical reaction to occur spontaneously, the overall difference
in free energy (G) between products and reactants must be negative.
4. Like all catalysts, enzymes merely accelerate the rate but do not
change the G of a reaction or the equilibrium between reactants and
products.
5. Enzymes reduce the activation energy of a reaction by providing an
alternative path from reactants to products, one that may break up the
reaction into smaller steps that are easier to overcome.
III.1. Michaelis-Menten equation

In this equation: V0 is the initial velocity of the reaction.

Vmax is the maximal rate of the reaction.
[Substrate] is the concentration of the substrate.
K is the Michaelis-Menten constant
Ex. KM (Michaelis constant) of some enzymes
 III.2 Lineweaver-Burk equation

This equation yields a straight line (y = mx + b), with y = 1/v, x = 1/[S],

m = slope, and b = y-intercept. Therefore a graph of 1/v versus 1/[S]
III.4. Eadie- Hofstee plot

In this case, a plot of v versus v/[S] has a y-axis (v-intercept) of

Vmax, an x-axis (v/[S]) intercept of Vmax/Km, and a slope of -
Km. The Eadie–Hofstee plot does not compress the data at high
substrate concentrations.
Enzyme kinetics plot. Kinetic representations of the properties
of enzymes.
(A) Michaelis–Menten plot of velocity (v) versus substrate
concentration ([S]). (B) Lineweaver–Burk plot. (C) Eadie–Hofstee
plot.
- Many external factors other than catalysts can affect the
rates of physiologic reactions.
1. In the absence of catalysis, a reaction can be
accelerated by adding energy in the form of heat, but this is
impractical in the body.
2. Increased concentration of one or more reactants also
accelerates a reaction by increasing occupancy of substrate
binding sites on available enzymes.
3. Enzymes normally operate within an optimal pH range
in which the important amino acids of the active site have
the correct state of protonation
Clinical test
Measurement of enzyme activity in
clinical samples.

Amylase Test
- Amylase blood and/or urine test if you have
symptoms of a pancreatic disorder.
 IV. MECHANISM OF ENZYME ACTION

- Enzymes use a variety of strategies to catalyze reactions, and

individual enzymes often use more than one strategy.
- Substrate binding by an enzyme helps catalyze the reaction by
bringing the reactants into proximity with the optimal orientation for
reaction.
- Amino acid side chains within active sites of many enzymes assist in
catalysis by acting as acids or bases in reaction with the substrate.
IV.1. Enzymatic reactions involve functional groups on
amino acid side chains, coenzymes, substrates, and
products.
- Enzymes vary significantly in their mechanism of
action. Such as: + catalysis is carried out on a substrate
noncovalently, reversibly bound to the enzyme.

+ a covalent intermediate is formed on and then

released from the enzyme.

+ all the action takes place on a coenzyme that

forms a covalent bond with substrate.
  The serine
proteases are
representative of
enzymes that form a
covalent intermediate
with their substrates.

A schematic model of a catalytic

triad of serine protease.
- Groups amino acid side chains participate in the enzyme-
catalyzed reaction. In the serine protease family, an
active-site serine residue catalyzes cleavage of the
peptide bond.
- Group OH (alcohol primary) is not among the more
reactive functional groups in organic chemistry.
- To enhance its activity in serine proteases, this serine
residue is part of a “catalytic triad,” in the case of
chymotrypsin: Asp(102), His(57), and Ser(195).
- In chymotrypsin cleavage on the carboxyl side of peptide
bonds containing aromatic amino acids, such as
phenylalanine. The mechanism of the enzymatic reaction
is outlined in Fig. bellow, showing the formation and
cleavage of an enzyme-bound intermediate.
Mechanism of action of chymotrypsin. The active-site serine residue attacks the carbonyl
group (-C(=O)- of the peptide bond on the carboxyl (-COOH) side of a phenylalanine residue.
The carboxy-terminal peptide is released, and the amino-terminal peptide remains an
enzyme-bound intermediate the amino-terminal peptide linked covalently through its
carboxy-terminal phenylalanine esterified to the active-site serine residue. The ester bond is
hydrolyzed in the second step of the reaction to release the amino-terminal peptide and
regenerate active enzyme.
IV.2 Enzyme Inhibitors
- Three forms of reversible inhibition: competitive,
uncompetitive, and noncompetitive inhibition.
- Enzyme inhibitors work in several ways and are clinically
important as drugs.
IV.2.1. Competitive inhibitors
Competitive inhibitors resemble the substrate in structure
and bind reversibly to the enzyme’s active site.
- Because a competitive inhibitor binds to the same site
on the enzyme as the substrate, it can be displaced by
increasing the substrate concentration, which overcomes the
inhibition.
- Competitive inhibitors increase the apparent Km while
having no effect on V max.
 Competitive enzyme inhibition. (A) Plot of velocity versus substrate
concentration. (B) Mechanism of competitive inhibition. (C) Lineweaver–
Burk plot in the presence of a competitive inhibitor. (D) Eadie–Hofstee
plot in the presence of a competitive inhibitor. K′m is the apparent Km in
the presence of inhibitor
IV.2.2. Uncompetitive inhibitors
Uncompetitive inhibitors cause an apparent decrease in
Vmax.
Questions:

1/ Why do uncompetitive inhibitors can cause an apparent

decrease in Vmax?
2/ Compare competitive inhibitors and uncompetitive inhibitors.
IV.2.3. Noncompetitive inhibitors

Noncompetitive inhibitors may bind to sites outside the active site

and alter both the Km and the Vmax of the enzyme.
IV.2.4. Irreversibly inhibit enzymes
Many drugs and poisons irreversibly inhibit enzymes

- Prostaglandins are key inflammatory mediators. Their synthesis is initiated by

cyclooxygenase-mediated oxidation and cyclization of arachidonate under inflammatory
conditions. Compounds that suppress cyclooxygenase have
anti-inflammatory activity. Aspirin (acetylsalicylic acid) inhibits cyclooxygenase
activity by acetylating Ser530, which blocks access of arachidonate to the active
site of the enzyme.
- Other nonsteroidal anti-
inflammatory drugs (NSAID), such
as indomethacin, inhibit
cyclooxygenase activity by
reversibly blocking the
arachidonate binding site.
Arachidonic acid metabolism
- Disulfiram (Antabuse) is a drug used for the treatment of alcoholism.
Alcohol is metabolized in two steps to acetic acid. The first enzyme,
alcohol dehydrogenase, yields acetaldehyde, which is then converted into
acetic acid by aldehyde dehydrogenase. The latter enzyme has an active-
site cysteine residue that is irreversibly modified by disulfiram, resulting
in accumulation of both alcohol and acetaldehyde in the blood. People
who take disulfiram become sick because of the accumulation of
acetaldehyde in blood and tissues, leading to alcohol avoidance.
- Alkylating agents, such as iodoacetamide (ICH2CONH2), irreversibly
inhibit the catalytic activity of some enzymes by modifying essential
cysteine residues. Heavy metals, such as mercury and lead salts, also
inhibit enzymes with active-site sulfhydryl residues. The mercury
adducts are often reversible by thiol compounds. Eggs or egg whites
are sometimes administered as an antidote for accidental ingestion of
heavy metals. The egg-white protein ovalbumin is rich in sulfhydryl
groups; it traps the free metal ions and prevents their absorption
from the gastrointestinal tract.
In many cases, irreversible inhibitors are used to
identify active-site residues involved in enzyme
catalysis and to gain insight into the mechanism of
enzyme action. By sequencing or mass spectrometric
analysis of the modified peptide, it is possible to
identify the specific amino acid residue modified by
the inhibitor and involved in catalysis.
V. Regulation of enzyme activity
There are multiple complementary mechanisms for
regulation of enzyme activity.
Generally, five independent mechanisms are involved in
the regulation of enzyme activity:
- The expression of the enzyme protein from the
corresponding gene changes in response to the cell’s
changing environment or metabolic demands.
- Enzymes may be irreversibly activated or inactivated
by proteolytic enzymes.
- Enzymes may be reversibly activated or inactivated by

covalent modification, such as phosphorylation.

- Allosteric regulation modulates the activity of key
enzymes through reversible binding of small molecules at
sites distinct from the active site in a process that is
relatively rapid and, hence, the first response of cells to
changing conditions.
Database bank of Allosteric molecules: https://mdl.shsmu.edu.cn/ASD/
Schematic representation of allosteric regulation with
positive cooperativity.
- The rate of degradation of enzymes by intracellular
proteases in the lysosome or by proteasomes in the cytosol
also determines the half-life of enzymes and, consequently,
enzyme activity over a much longer period of time.
Feedback inhibition
Clinical Box
Hemophilia is caused by a defect in
Zymogen activation
A child was admitted to hospital with muscle bleeding
affecting the femoral nerve. Laboratory findings indicated
a blood-clotting disorder, hemophilia A, resulting from
deficiency of factor VIII. Factor VIII was administered to
the patient to restore blood-clotting activity.
 Clinical Box
Insecticide poisoning

A 55-year-old man was spraying an insecticide

containing organic fluorophosphates in a rice field. He
suddenly developed a frontal headache, eye pain, and
tightness in his chest, typical signs of overexposure to
toxic organic fluorophosphates. He was taken to the
hospital and treated with an intravenous injection of 2 mg
of atropine sulfate, and he gradually recovered.
Clinical Box
Insecticide poisoning

Comment

Organic fluorophosphates form covalent phosphoryl-enzyme

complexes with both serine proteases and esterases, such as
acetylcholinesterase, irreversibly inhibiting the enzymes.
Acetylcholinesterase terminates the action of acetylcholine during
neuromuscular activity by hydrolyzing the acetylcholine to acetate
and choline. Inhibition of this enzyme prolongs the action of
acetylcholine, leading to constant neuromuscular stimulation.
Atropine competitively blocks acetylcholine binding and muscle
stimulation at the neuromuscular junction.
Clinical Box
Insecticide poisoning

Diisopropyl fluorophosphate (DIFP)

Irreversible inactivation of a serine protease by the suicide inhibitor DIFP

(Diisopropyl fluorophosphate, isofluorophate).
LABORATORY TESTING
ENZYMATIC MEASUREMENT OF BLOOD GLUCOSE
The glucose oxidase/peroxidase assay
In clinical laboratories, most compounds are measured by automated enzymatic methods.

The most common assay for blood glucose concentration uses a mixture of glucose oxidase and
peroxidase (Fig above). Glucose oxidase is highly specific for glucose but oxidizes only the β-
anomer of the sugar, which represents ~64% of glucose in solution. The assay mixture is therefore
supplemented with mutarotase, which rapidly catalyzes the interconversion of the anomers,
enhancing assay sensitivity by ~50%. The H2O2 produced in the oxidase reaction is then used in a
peroxidase reaction to oxidize a chromogen to yield a colored chromophore. The color yield is
directly proportional to the glucose content of the sample. There are fluorometric versions of this
assay for high sensitivity, and one commercial analyzer uses an oxygen electrode to measure the
rate of decrease in oxygen concentration in the sample, which is also directly proportional to the
glucose concentration .
LABORATORY TESTING
ENZYMATIC MEASUREMENT OF BLOOD GLUCOSE

Reagent strips and glucometers

Kinetic assays
Kinetic assays are more rapid than endpoint assays
LABORATORY TESTING
ENZYMATIC MEASUREMENT OF BLOOD GLUCOSE

Kinetic analyzers are inherently faster than endpoint assays

because they estimate glucose concentration before the assay reaches
its endpoint. These assays work because glucose oxidase and glucose
dehydrogenase have a high Km for glucose. At the concentrations of
glucose found in blood, the rate of the oxidase reaction is proportional
to glucose concentration - that is, in the first-order region of the
Michaelis–Menten equation, where the substrate concentration is less
than the Km.
CHAPTER 4: VITAMINS AND MINERALS
 LEARNING OBJECTIVES
LEARNING OBJECTIVES

- Describe fat-soluble and water-soluble

vitamins.
- Describe the actions and sources of vitamins.
- Discuss signs and symptoms of vitamin
deficiencies
- Describe the role of trace metals in
metabolism.
 I. Introduction
- Vitamins are organic compounds required by the body
in small amounts for metabolism, for protection, for
maintenance of health and proper growth.
- They cannot be synthesized by the body. Must be
obtained by outside sources like diet, rumen of
bacteria & sun.
- Vitamins also assist in the formation of hormones,
blood vessels, nervous system chemicals and genetic
materials.
- They generally act as catalysts, combining with
proteins to create metabolically active enzymes that
are essential for life reactions. Without enzymes,
many of the reactions essential to life would slow
down or cease.
II. CLASSIFICATION OF VITAMINS
- Two main groups of vitamins: fat-soluble
vitamins and water-soluble vitamins. The body
can store fat-soluble vitamins, but any excess
water-soluble vitamins are easily removed from
the body in the urine, so regular intake is
necessary.
 Table: Vitamins essential for human health, reference nutrient intake (RNI)
values taken from the UK Food Standards Agency website, and the main
dietary sources of these vitamins.

Name RNI values for adults per Main dietary sources

day
Fat-soluble vitamins
vitamin A 0.6 mg for women; 0.7 liver, cheese, eggs, butter, oily fish
mg for men (such as mackerel), milk, yoghurt

vitamin D 0.01 mg (10 μg) for oily fish, liver, eggs, margarine,
certain groups, e.g. bread, powdered milk
pregnant women.
vitamin E 3 mg for women; 4 mg plant oils (such as soya, corn and
for men olive oil), nuts, seeds, wheat
germ, some green leafy vegetables
vitamin K 0.07 mg(70 μg), or 1 μg green leafy vegetables (such as
per kg of body weight broccoli and spinach), vegetable
oils, cereals; small amounts can
also be found in meat (such as
pork), and dairy foods (such as
cheese)
 Name RNI values for adults Main dietary sources
per day
Water-soluble vitamins
thiamin (vitamin 0.8 mg for women; 1 pork, vegetables, milk, cheese,
B1 ) mg for men peas, fresh and dried fruit,
eggs...
riboflavin (vitamin 1.1 mg for women; 1.3 milk, eggs, rice, mushrooms.
B2 ) mg for men

niacin (vitamin B3 ) 13 mg for women; 17 beef, pork, chicken, wheat flour,

mg for men maize flour, eggs, milk

vitamin 1.2 mg for women; 1.4 liver, pork, chicken, cod, bread,
B6 (pyridoxine) mg for men whole cereals (such as oatmeal,
wheatgerm and rice), eggs,
vegetables, soyabeans, peanuts,
milk, potatoes
folate (folic acid, 0.2 mg, but 0.4 mg broccoli, sprouts, spinach, peas,
vitamin B9) extra for women who chickpeas, potatoes, yeast
are, or plan to be, extract, brown rice, some fruit
pregnant (such as oranges and
bananas),breakfast cereals,
 Name RNI values for adults Main dietary sources
per day
Fat-soluble vitamins
vitamin 0.0015 mg (1.5 μg) meat (particularly liver),
B12 (cobalamin) salmon, cod, milk, cheese, eggs,
yeast extract
pantothenic acid none given – should be chicken, beef, potatoes,
(vitamin B5) sufficient in normal porridge, tomatoes, liver,
diet kidneys, eggs, broccoli,
wholegrains (such as brown rice
and wholemeal bread),...

biotin (vitamin 0.01–0.2 mg meat (such as kidney and liver),

H) eggs and some fruit and
vegetables, especially dried
mixed fruit
vitamin C 40 mg wide variety of fruit and
(ascorbic acid) vegetables, especially peppers,
broccoli, sprouts, sweet
potatoes, blueberries, citrus
fruits, kiwi fruit
 FAT SOLUBLE VITAMINS

1/ VITAMIN A: Vitamin A is a
pale yellow primary alcohol
derived from carotene.
- It includes Retinol (alcoholic
form), Retinal (aldehyde form)
and Retinoic acid (acidic form).
- Vitamin A deficiency cause
blindness,
anaemia, conjunctivitis
Xerophthalmia /dry-eye;
Children who are vitamin A-
deficient are more susceptible
to respiratory infections and
measles, cardiovascular
diseases, diabetes mellitus,
obesity, osteoporosis, skin
diseases and cancer ...
A schematic representation of the physiological roles in which vitamin A
is involved
Vision and the role of 11-cis-retinal in the process. The retina comprises
cones and rods, which mediate color and low light vision, respectively. The
vitamin A derivative 11-cis retinal is found in the rods, forming rhodopsin.
QUESTION

Why is 11-cis – Retinal? Why not 7-cis-, 9-cis-, or 13-

cis- Retinal in the Eye?
2/ Vitamin D: This comprises a group of fat-soluble
sterol founds naturally in few foods.
- The two major physiologically relevant forms of
vitamin D are D2 (ergocalciferol) and D3
(cholecalciferol).
The synthesis of vitamin D3 (cholecalciferol, D3) occurs at the skin where pro-vitamin D3
(7-dehydrocholesterol) is converted to pre-vitamin D3 in response to sunlight exposure
(ultraviolet B radiation). Vitamin D3, obtained from the isomerization of pre-vitamin D3 in
the epidermal basal layers, or intestinal absorption of natural and fortified foods and
supplements D2 (ergocalciferol) and D3, binds to vitamin D-binding protein (DBP) in the
bloodstream, and is transported to the liver. D2 and D3 are hydroxylated by liver 25-
hydroxylases. The resultant 25-hydroxycholecalciferol [25(OH)D] (calcifediol or calcidiol) is
1-hydroxylated in the kidney by 1α-hydroxylase. This yields the active secosteroid
1,25(OH)2D (calcitriol), which has different effects on various target tissues. The synthesis of
1,25(OH)2D from 25(OH)D is stimulated by the parathyroid hormone and suppressed by
calcium, phosphate, and 1,25(OH)2D itself.
Biological actions of Vitamin D:
Calcium and phosphorus homeostasis:
- Increased intestinal calcium absorption and synthesis of
the intestinal calcium transporter;
- Increased intestinal absorption of phosphorus;
- Increased renal reabsorption of calcium and
phosphorus;
- Induction of mature osteoblasts differentiation from
precursors;
- Stimulation of bone resorption.
Immunomodulatory effects
- Induction of monocytes differentiation to
macrophages;
- Increased rate of phagocytosis;
- Increased production of lysosomal enzymes;
- Decreased interleukin-2 production;
- Increased interleukin-10 production.
Antitumor effect:
- Induction of cell differentiation;
- Increased apoptosis of neoplastic cells.
Cardiovascular effect:
- Reduction of plasma renin activity and angiotensin II
3/ VITAMIN E: also called Tocopherol or fertility
hormone Vitamin E is required in the human diet but
its deficiency is rare except in pregnancy and the
new born, where it is associated with hemolytic
anaemia.
• It exists in the diet as a mixture of eight closely
related compounds called tocopherols.
4/ VITAMIN K: Also called
phylloquinone or anti-hemorragic
vitamin or coagulation vitamin.
- Vitamin K is a complex
unsaturated hydrocarbon found in
two forms, vitamin
K1(phylloquinone) and vitamin
K2(Menaquinone)
- Vitamin K deficiency is not
common in adults, the symptoms
of vitamin K deficiency are
spontaneous cutaneous purpura,
epistaxis, gastrointestinal,
genitourinary, gingival, or other
bleeding .
Structures of the different forms of vitamin K
 WATER SOLUBLE VITAMINS

- These include the group B-vitamins and vitamin C

- They are soluble in water and can therefore be

excreted in the urine. They share few common
properties besides their solubility characteristics.

- Most of these vitamins act as coenzymes. Examples

include thiamine (Vit B1), Riboflavin (B2), Niacin,
Pantothenic acid (Vit B5), Vitamin B6 (Pyridoxine),
Biotin, Vitamin B12 (Cobalamin) & folic acid.
VITAMIN B1(Thiamine):
- It serves as a cofactor for several enzymes involved in
energy metabolism.
- Important: biosynthesis of neurotransmitters and reducing
substances used in oxidant stress, synthesis of pentoses
used as nucleic acid precursors.
- To play a central role in cerebral metabolism.
- It is a colorless and crystalline substance.
- It is readily soluble in water and slightly in ethyl alcohol.
- Addition of a pyrophosphate (ppi) from ATP converts it to
thiamine pyrophosphate (TPP), the coenzyme for all
decarboxylation of alpha keto acids.
- B1 deficiency results in dry/
wet beriberi, a peripheral
neuropathy, a cardiomyopathy
with edema and lactic acidosis,
Wernicke–Korsakoff syndrome,
ophthalmoplegia, and ataxia
evolving into confusion,
retrograde amnesia, cognitive
impairment, and confabulation.
 B1 vitamin is a cofactor
required for several metabolic
processes.
  The three
thiamine-dependent
enzymes and their
role in the
pathogenesis of cell
death in thiamine
deficiency.
VITAMIN B2 (Riboflavin):

- It is required for FMN and FAD synthesis

- It is composed of an isoalloxazane ring system linked to ribitol

- It is mainly used in energy metabolism of sugars and lipids

- The activation of FMN and FAD is an ATP-dependent reaction

Laboratory test

To determine riboflavin status, erythrocyte glutathione

reductase activity is measured.
Vitamin B3 (Niacin):
- Niacin can be synthesized from tryptophan.
- Niacin contains a substituted pyridine ring and when
NAD+ activated forms NAD+ and its phosphorylated
derivative NADP+, which are coenzymes of many
dehydrogenases.
VITAMIN B5 (pantothenic acid)
- Pantothenic acid is widely distributed in animals and Plants
Pantothenic acid forms part of the molecule of coenzyme A
(CoA ).
- There is no evidence of deficiency in humans, except those on
experimental diets.
VITAMIN B6 (PYRIDOXINE)
- Vitamin B6 (pyridoxine) participates in carbohydrate
and lipid metabolism and is particularly important for
amino acid metabolism

Vitamin B6 is a mixture of pyridoxine, pyridoxal,

pyridoxamine, and their 5′-phosphate esters.

- Pyridoxine requirements increase with high protein

intake

- Pyridoxine deficiency causes neurologic symptoms

and anemia
 Vitamin B6 deficiency in its mild form causes irritability,

nervousness, and depression, progressing in severe

deficiency to peripheral neuropathy, convulsions, and coma.

Severe deficiency is also associated with a sideroblastic

anemia (anemia characterized by the presence of nucleated

red blood cells with iron granules). Dermatitis, cheilosis, and

glossitis also occur.

- Assessment of pyridoxine status is based on the

measurement of erythrocyte aspartate aminotransferase.

VITAMIN B7 (BIOTIN)
- Biotin is a vitamin and a coenzyme commonly
associated with enzyme performing carboxylation
reactions.
- Biotin is also known as “anti-egg white injury factor”
or as H-factor.
VITAMIN B9 or M or Bc (folic acid)
- Vitamin B9 (folic acid) derivatives are important in single-carbon-
transfer reactions and are necessary for the synthesis of DNA.

- Structural analogues of folate are used as antibiotics and

anticancer drugs.

- Folate deficiency is one of the commonest vitamin deficiencies

- Folate deficiency in adults causes megaloblastic anemia

- Adequate intake of folate around conception is essential

VITAMIN B12 (Cobalamin)
- Vitamin B12 forms part of the heme structure

- Vitamin B12 requires the intrinsic factor

(glycoprotein) for its absorption

- Vitamin B12 is only present in animal

products

- Vitamin B12 deficiency causes pernicious

anemia: Vitamin B12 deficiency is characterized
by anemia, fatigue, constipation, weight loss,
diarrhea, and neurologic symptoms such as
numbness and tingling, loss of balance,
confusion, mood disturbances, and dementia.
ASCORBIC ACID, VITAMIN C

- It is a water soluble vitamin

- Vitamin C takes part in the synthesis of collagen and

epinephrine, in steroidogenesis, in the degradation of tyrosine,
in the formation of bile acids, and also in the synthesis of L-
carnitine and neurotransmitters. It improves absorption of
nonheme iron and participates in bone mineral metabolism.

- Its prime function is to maintain metal cofactors in their lower

valence states (e.g., Fe2+ and Cu2+). In the synthesis of
collagen, it is required specifically for the hydroxylation of
proline.
Proline and lysine hydroxylases are
necessary for the post-synthetic
conversion of procollagen to collagen
 Vitamin C is absorbed in the intestine by a carrier-mediated,
sodium-dependent transporter. It is reabsorbed in the renal
proximal tubules. Progressively more vitamin C is excreted in urine
as intake increases.

- Humans cannot synthesize ascorbic acid; therefore it is an

essential nutrient.

- Vitamin C deficiency causes scurvy and compromises immune

function.
No clear evidence exists, however, to substantiate these
claims first made by Linus Pauling in the 1970s. Vitamin C is
certainly required for normal leukocyte function, and leukocyte
vitamin C levels drop precipitously during the stress caused
by either trauma or infection. .
Structure, sources, and deficiency diseases of the B vitamins
Summary
Role of vitamins in metabolism
However,
- The benefits of vitamin supplementation in cancer and
cardiovascular disease are uncertain.
- Vitamin supplementation can be harmful
- Fruit and vegetables are the best sources of vitamins
 MINERALS
- Minerals are inorganic elements needed for the
functioning of the body
- They make up about 4% of body weight of adults,
they cannot be changed or broken down
- Some which are needed in high quantities are
referred to as macro-elements, examples include
Na, K, Mg, Ca etc.
- Others are needed in smaller quantities and are
termed microelements, they include Fe, Cu, F, I
etc.
The main elements that compose the human body
- The body requires relatively large amounts of
about 7 minerals (macro-elements)
1/ Calcium (Ca)
2/ Phosphorus (P)
3/ Sulphur
4/ Magnesium: Magnesium (Mg): it acts as coenzymes
for enzymes
5/ Potassium
6/ Chlorine
7/ Sodium
The body requires only trace amounts of others
(micro-elements). These includes:
1/ Flouride (F)
2/ Iodine (I)
3/ Iron (Fe) (See Chapter: Iron metabolism).
4/ Cobalt (Co)
5/ Chromium (Crom)
6/ Manganese
7/ Copper (See Chapter: Copper metabolism)
8/ Zinc (See Chapter: Zinc metabolism)
9/ Molybdenum (Mo)
10/ Selenium
....
Selenium
Case report of selenium responsive cardiomyopahty
(Abdulrahman Al-Matary al et., 2013)

The authors report a case of selenium-responsive cardiomyopathy in

a 15-month old Saudi Arabian boy. This case of selenium deficiency
causing dilated cardiomyopathy is presented with failure to thrive,
prolonged fever and respiratory distress. The investigations revealed
selenium deficiency. Selenium supplementation along with anti-failure
therapy [Furosimide, Captopril] was administered for 6 months. Following
therapy the cardiac function, hair, skin and the general health of the
patient improved significantly.

What are the signs of selenium deficiency?

(Selenium: health benefits, sources and potential risks)

Selenium

- Selenium is present in all cells as amino acids selenomethionine

and selenocysteine

- Selenium status may influence the risk of many chronic conditions

There is a rare selenium-responsive cardiomyopathy (Keshan

disease), which is endemic in China in areas of very low selenium
intake. Selenium deficiency may result in chronic muscle pain,
abnormal nail beds, and cardiomyopathy. Excess of selenium, on the
other hand, leads to liver cirrhosis, splenomegaly, gastrointestinal
bleeding, and depression.
Selenium
 Selenium

Se, selenium; Sec, selenocysteine; HSe–,

hydrogen selenide; SEPHS2,
selenophosphate synthetase 2; GPX,
glutathione peroxidase; TXNRDs, thioredoxin
reductases; DIO, iodothyronine deiodinases;
SELENOP, selenoprotein P; SELENOT,
selenoprotein T; SELENOK, selenoprotein K;
MSRB1, methionine sulfoxide reductase B1;
SELENOW, selenoprotein W
Biochemical Endocrinology
LEARNING OBJECTIVES
LEARNING OBJECTIVES
- 1/ Give the definition of a hormone, and explain its action in
endocrine, paracrine, and autocrine systems.
- 2/ Explain the classification of hormones according to their
structure.
- 3/ Describe the organization and regulatory role of the
hypothalamus and pituitary.
- 4/ Explain the processes controlling the biosynthesis, transport,
and action of thyroid hormones.
- 5/ Explain the mechanisms regulating the synthesis and action
of glucocorticoids.
- 6/ Describe the regulation of the synthesis and activity of sex
steroid hormones and their role in human reproduction.
- 7/ Explain the direct and indirect actions of growth hormone.
- 8/ Describe the actions of prolactin.
- 9/ Describe the clinical manifestations of deficiencies and
excesses of hormones.
- 10/ Describe the principles of investigating endocrine
dysfunction.
I. Introduction:
- To regulate physiological activities and in
maintaining homeostasis.
- Specific organs or tissues
- To diffuse across cell membranes and discharge
bloodstream after they are secreted from endocrine
glands.
- There are endocrine, paracrine, and autocrine
hormones.
1. Endocrine hormones (e.g., cortisol, insulin, and
prolactin) are those messengers transported in the
blood to a target distant from the site of secretion.
2. Paracrine hormones (e.g., neurotransmitters and
growth
factors) exert their action locally at the site of
secretion.
3. Autocrine hormones (e.g., IL-2 in activated
lymphocytes) when hormones act on the cells of their
synthetic origin
These can influence their own hormonogenesis.
Relationships between endocrine and neural regulation
II. Classification of hormones

Structurally, hormones may be modified amino acids,

peptides, glycoproteins, or steroids.

(1) modified amino acids

(2) peptides

(3) glycoproteins
(4) steroids.
Chemical derivation of hormones
III. Principles of hormone action

Endocrine systems exhibit core general properties: (1)

release of hormones in response to a stimulus, (2) transport of
the hormone to the target tissue, (3) hormone stimulation of cell
receptors, (4) feedback regulation of hormone secretion (Fig.
bellow), and (5) clearance of hormones.

Basic endocrine processes.

 IV. Regulation of hormone production
Hormone systems are typically controlled by feedback
mechanisms
- Negative feedback: the most common form. An example is the effect of
thyroxine (T4) and triiodothyronine (T3) on the hypothalamus and
pituitary. Negative feedback mechanisms do not exist without external
influence; if they did, then hormonogenesis would remain constant. Indeed,
many endocrine organs, particularly those that are under hypothalamic
control, exhibit rhythmicity, which is influenced by neuronal input.

- Positive feedback: This is rarer, and examples include the secretion of

LH in the female menstrual cycle, where the hormone concentrations
quickly increase before ovulation.
Hormone degradation and clearance

The inactivation of hormones is key to their function as

controllers of homeostasis

+ Decreased secretion

+ Hormone degradation. Hormone degradation may occur

in the blood, in organs such as the liver or kidneys, or in
target tissue itself. The clearance of hormones varies widely,
from minutes (insulin), to hours (glucocorticoids), to days
(T4). Clearance can also alter in disease states, such as the
delayed clearance of insulin that may be observed in liver
disease.
v. Causes of endocrine disease
- Autoimmunity and neoplasia
- Exogenous hormone administration

VI. The hypothalamus and the pituitary gland

 PIH

PRH

Hypothalamo–anterior–pituitary regulatory target organ

axes
Adenohypophysis: 7 hormones.

1/ Growth hormone (somatotropin) (GH)

2/ Prolactin

3/ Adrenocorticotropic hormone (ACTH)

4/ Thyrotropin (thyroid-stimulating hormone) (TSH)

5/ Follicle-stimulating hormone (FSH)

6/ Luteinizing hormone (interstitial-cell-stimulating

hormone) (LH)

7/ Melanocyte-stimulating hormone (intermedin) (MSH)

GH hormone disorders

1/ Dwarfism
2/ Gigantism
3/ Acromegaly

Gigantism
Posterior pituitary
1/ Oxytocin

Oxytocin stimulates smooth muscle contraction in the uterus

and breast, functioning in childbirth and breastfeeding,
respectively. It is released in response to stimulation of
mechanoreceptors in the breast with suckling. Synthetic oxytocin
may be used to induce labor or to control uterine bleeding after
childbirth.

2/ Vasopressin ( or Antidiuretic Hormone: ADH)

They are two peptide hormones synthesized in the cell

bodies of the hypothalamic neurons that are subsequently
secreted by the posterior pituitary.
Hormones of the thyroid gland
1/ Triiodothyronine (T3)
2/ Thyroxine (T4)
Hormones of the pancreas
1/ Insulin
2/ Glucagon
Hormones of the adrenal glands.
Clinical box
Acute glucocorticoid withdrawal

Case:
A 47-year-old man presented to the emergency department with
persistent nausea, vomiting, lethargy, and generalized abdominal
pain after a bout of food poisoning. He had been able to drink some
fluids but had been unable to keep food or his tablets down. He had
a history of chronic severe asthma that was recently well controlled
with inhalers and long-term oral glucocorticoids.
On examination, there was a mild bilateral wheeze in his lungs;
his abdomen was soft and nontender, and bowel sounds were
present. His blood pressure was 115/65 mmHg lying down. Venous
blood glucose was 3.8 mmol/L 68 mg/dL (4–6 mmol/L; 72–109
mg/dL). The patient was administered hydrocortisone and fluids
intravenously and made a full recovery.
Clinical box
Acute glucocorticoid withdrawal

Comment:

This presentation is an acute hypoadrenalism following sudden

withdrawal of glucocorticoid therapy. Due to long-term exogenous
glucocorticoid use, patients can have adrenal atrophy (due to lack of
ACTH production). Stress can precipitate an adrenal crisis in patients
who are unable to mount an adequate cortisol response, and “sick-
day rules” are taught so that individuals can increase their dose of
steroids during periods of illness.
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ATTENTION!