218

Protein
Metabolism
Objectives
 Explain digestion of proteins
 Explain nitrogen balance
 Differentiate and explain essential, non essential,
glucogenic and ketogenic amino acids
 Comment on the role of transamination and oxidative
deamination
 Explain ammonia intoxication or hyperammonemia
 Explain synthesis of urea and disorders of urea cycle
enzymes
 List the plasma protein and their functions
 Comment on the role of hormones on protein
metabolism
 List and identify the clinically significant NPN
compounds
 Types of nitrogen balance
Nitrogen equilibrium : Intake = excretion In healthy individuals

 Positive N balance  Negative N balance

 Intake> excretion
 During infancy ,
childhood, adolescence
 Pregnancy
 Lactation
 Recovery after
surgery,burns,trauma,s
tress
 Intake < excretion
 Inadequate calorie and
protein intake
 starvation , infection ,
prolonged immobility
and stress
 Immediately after
surgery and crush
injuries
Digestion of dietary proteins
 Most nitrogen in the diet is consumed in the form
of protein
 Proteins are too large to be absorbed by the
intestines
 They must be hydrolyzed to yield constituent
amino acids which can be absorbed
 Breakdown of proteins provide amino acids as fuels
 When amino acids are metabolized their nitrogen
must be disposed of in non toxic form
 Enzymes responsible for degrading proteins are
produced by (1) stomach,(2) pancreas and (3)
small intestine
Cont…
Dietary protein
↓pepsin
polypeptides and amino acids
↓elastase,trypsin,chymotrypsin,carboxypeptidases
Oligopeptide and amino acid
↓aminopeptidases
Free Amino acids and dipeptides
Cont…

stomach pancreas to intestinal

small intestine wall
pepsin Trypsin dipeptidases

Chymotrypsin

carboxypeptidase A

carboxypeptidase B

elastase
 OVERVIEW OF AMINO ACID METABOLISM

ENVIRONMENT ORGANISM

Bio-
Ingested synthesis
Protein
protein

AMINO
ACIDS

Degradatio Purines
n Pyrimidines
(required) Porphyrins
Carbon
Nitrogen
skeletons
(ketogenic) (glucogenic)
Urea Used for
pyruvate
energy α-ketoglutarate
acetoacetate
acetyl CoA succinyl-CoA
fumarate
oxaloacetate
First phase of amino acid metabolism
Removal of α-NH2 groups by transamination and subsequent
oxidative deamination leads to formation of ammonia and
corresponding keto acid (carbon skeleton)
Small amount of ammonia is excreted in urine but most is
used in synthesis of urea
Synthesis of urea is the most important route for disposing
of nitrogen from the body
Second phase of amino acid metabolism
The carbon skeletons are converted to common
intermediates of energy producing metabolic pathways
These intermediates can be metabolized to CO2,H2O,Fatty
acid or ketone bodies
Third phase of amino acid metabolism
Biosynthesis of amino acids
Biosynthesis of amino acid derivatives like
GABA,catecholamines,creatine,histamine,melanins,S-
adenosylmethionine etc
Removal of NH2 group and Transamination
 Presence of α- NH2 group keeps the amino acids safely locked from
OXIDATIVE DEAMINATION
 Removal of α- NH2 is essential for producing energy from amino acid and
is done by TRANSAMINATION
 TRANSAMINATION and OXIDATIVE DEAMINATION are
reactions that remove NH2 group from amino acids
 AND ultimately provide ammonia and aspartate, which are two sources of
urea nitrogen
 Transamination is defined as transfer of amino group from an amino
acid to a keto acid to form another ( new) amino acid and keto acid of the
original amino donor
 Most amino acids undergo transamination EXCEPT
lysine,threonine,proline and hydroxyproline
 Enzymes responsible for transamination are known as transaminases
Cont…
Transaminases can function in both amino acid catabolism and
synthesis
α-NH2 groups of amino acids are ultimately transferred by transaminases
to form L- glutamate. PLP acts as coenzyme in transamination
reactions.
This nitrogen is released as ammonia (NH3) by a reaction catalysed by
glutamate dehydrogenase that uses NAD+ and NADP+ as coenzymes
ALT- Alanine amino transferase catalyses transfer of amino group to
form pyruvate and glutamate
AST- Aspartate amino transferase catalyses transfer of amino group to
form oxaloacetate and glutamate
COO COO COO COO
CH2 CH2 COO CH2 COO CH2
CH3 CH2 CH3 CH2 CH2 CH2 CH2 CH2
HC NH3+ + C O C O + HC NH3+ NH3+ + NH3+
HC C O C O + HC
COO COO COO COO COO COO COO COO
alanine -ketoglutarate pyruvate glutamate aspartate -ketoglutarate oxaloacetate glutamate
Aminotransferase (Transaminase) Aminotransferase (Transaminase)
Diagnostic value of amino transferases
 Are intracellular enzymes with low
levels found in plasma
 Presence of elevated levels indicate
damage to cells rich in these
enzymes
 AST and ALT are elevated in all liver
diseases
 ALT is more specific for liver
disease
 AST is more sensitive because liver
contains more amount of AST. AST
also increases during MI
 Oxidative Deamination
 Net conversion of α-NH2 to NH3
 Glutamate is the only amino acid that undergoes
oxidative deamination catalyzed by Glutamate
dehydrogenase and regenerates α-keto glutarate.

 The α-ketoglutarate formed can be used in citric

acid cycle and glucose synthesis
 Oxidative deamination requires the combined
effort of transaminases and glutamate
dehydrogenases and is also referred to as
transdeamination
 Regulation of glutamate dehydrogenase
(GDH)
 Glutamate dehydrogenase is
allosterically inhibited by ATP and GTP

 Thus when energy levels are low, AA

degradation by GDH is high

 It is reversible reaction and functions

both in AA synthesis and catabolism.
 Ammonia Intoxication or hyperammonemia

 High ammonia would deplete glutamate – a

neurotransmitter & precursor for synthesis of the
neurotransmitter GABA.
 Depletion of glutamate & high ammonia level
would drive Glutamate Dehydrogenase reaction
to reverse:
glutamate + NAD(P)+    a-ketoglutarate +
                                                             NAD(P)H + NH4+ 
 The resulting depletion of a-ketoglutarate, an
essential Krebs Cycle intermediate, could impair
energy metabolism in the brain
Cont…
 Symptoms of ammonia intoxication are
tremors,slurred speech,blurred vision, coma
and ultimately death

TREATMENT
 limiting protein intake to the amount barely
adequate to supply amino acids for growth,
while adding to the diet the a-keto acid
analogs.
  Liver transplantation has also been used,
since liver is the organ that carries out Urea
Cycle.
Overall flow of nitrogen in
amino acid catabolism
Fate of Ammonia produced by the tissues
 Ammonia produced by tissues is rapidly removed from
circulation by the liver and converted to
1. Glutamate
2. Glutamine
3. Finally to UREA
 Fate of Ammonia
 Ammonia produced by tissues is Glutamine transports ammonia in
rapidly removed from circulation blood stream
by the liver and converted to The amide of glutamine provides
a non toxic storage and transport
1. Glutamate
form of ammonia
2. Glutamine Glutamine synthesis is catalyzed
3. Finally to UREA by glutamine synthase
Glucose-Alanine Cycle
 Alanine transports
amino groups from
muscle to liver in non
toxic form via
pathway called as
glucose alanine cycle
 In muscle amino
groups are collected
as glutamate
 Glutamate transfers
its amino group to
pyruvate (obtained
from muslce
glycolysis) in presence
of ALT to form alanine
Glucose Alanine cycle
 Alanine passes into the
blood and travels to the
liver
 Liver ALT transfers
amino group from
alanine to α-KG to form
pyruvate and Glutamate
 Glutamate enters
mitichondria and is
acted upon by GDH to
release ammonia
 Pyruvate by
gluconeogenesis is
converted to glucose .
Liver glucose travels
through blood to muscle
 Urea cycle
 The urea cycle (also known as the ornithine cycle

 The Urea Cycle occurs mainly in liver

 Urea passes into the kidneys via the bloodstream and is finally
excreted in urine.
 The first two reactions of urea cycle occurs in mitochondrial
matrix while the next three reactions occur in the cytosol
 The 2 nitrogen atoms of urea enter the Urea Cycle as NH3
(produced mainly via Glutamate Dehydrogenase) and as the amino
N of aspartate.
 Synthesis of 1 molecule of urea requires 3 molecules of ATP plus 1
molecule each of ammonium ion and of α amino nitrogen of
aspartate
 Carbamoyl Phospahte Syntahase I (CPS I) is the rate limiting
enzyme and is activated by N-acetylglutamate
UREA CYCLE
Fate of Urea
 Urea diffuses from the liver and is transported in
blood to the kidneys, where it is excreted in the
urine
 Small amount of urea diffuses from the blood to
intestine where it is cleaved to CO2 and NH3 by
bacterial urease
 In patients with kidney failure plasma urea
levels are elevated greater transfer of urea
from blood to gut . The intestinal action of urease on
this urea Hyperammonemia
Urea cycle disorders
1. Hyperammonemia type 1. Is caused due to
the deficiency of CPS I
2. Hyperammonemia type 2. is caused due to
ornithine transcarbamoylase deficiency.
3. Citrullinemia : is caused due to the
deficiency of argininosuccinate synthase .
4. Argininosucciniaciduria ; is caused due to
absence of argininosuccinase
5. Hyperargininemia ; Is characterized by low
erythrocyte levels of arginase
Fate of carbon skeletons of Amino Acids
 Carbon skeletons of amino acids are converted
to SEVEN molecules which are either
intermediates of TCA cycle or substrates of
TCA cycle
 From here the carbon skeletons are diverted
to gluconeogenesis or ketogenesis or are
completely oxidised to CO2 and H2O
 The products formed are (1) OAA, (2) α KG,
(3) pyruvate, (4) fumarate (5) Succinyl CoA,
(6) Acetyl CoA,(7) Acetacetyl CoA
 Amino acids that form 1- 5 are called
Glucogenic and and which form 6-7 are
called ketogenic amino acids
Glucogenic and ketogenic amino acids
Glucogenic and ketogenic amino acids

 Amino acids, tryptophan, phenylalanine,

tyrosine, isoleucine and threonine are both
glucogenic and ketogenic.
 Only 2 amino acids are purely ketogenic they
are lysine and leucine.
 Amino acids that are purely glucogenic: Arg,
Glu, Gln, His, Pro, Val, Met, Asp, Asn,Ala,
Ser, Cys, and Gly.
Biosynthesis Of Amino Acids
 Humans can
synthesize 12 amino
acids from glycolysis
and TCA cycle
intermediates. These
amino acids are
nutritionally non
essential
 Nutritionally
Essential amino acids
cannot be synthesized
in the human body
and have to be
provided in the diet
 Biosynthesis of amino
acids can be divided
into six families
 Biosynthesis of Amino acids
II. PEP + ERYTHROSE-4-Phosphate
I. OAA

Aspartate LYS
Tyr
MET PHE TRP
Asparagine THR
Tyr
ILE
Biosynthesis of Amino acids

IV. Ribose-5-Phosphate

III. PYRUVATE

Ala VAL LEU

HIS
 Biosynthesis of Amino acids

V. α Ketoglutarate VI. 3-Phosphoglycerate

Glutamate
Serine

Arginine Glycine
Glutamine Cysteine
Pro
Conversion of Amino acids
to specialized products
 GABA-is synthesized from glutamate
 Histamine from histidine
Catecholamines- are synthesized from tyrosine
Serotonin is synthesized from tryptophan
Hormonal Regulation

 Insulin
 Glucose availability to cells increases
 Protein synthesis increases
 Glucagon
 Protein synthesis decreases
 Protein degradation increases
Plasma Proteins: Classification
1- Simple proteins
The simple proteins are those which are made of
amino acid units only, joined by peptide bond. Upon
hydrolysis they yield mixture of amino acids and
nothing else
e.g. albumin and globulin
2- Conjugated proteins
Conjugated proteins are composed of simple proteins
combined with a non-proteinous substance
Protein + prosthetic group.
e.g. lipoprotein,phosphoprotein,hemoglobim
3- Derived proteins
These are not naturally occurring proteins and are
obtained from simple proteins by the action of
enzymes and chemical agents e.g. peptones
 Plasma Protein Distribution

Fibrinogen
(4%) (1%)Other PlasmaProteins Albumin (60%)
Globulin (35%)
Globulin (35%)

Globulin Fibrinogen (4%)

Albumin
(35%)

(60%)
Other Plasma
Proteins (1%)
Other Plasma Proteins

 remaining one 1% of plasma

 Peptide hormones
 Insulin
 Prolactin
 Glycoproteins
 TSH (thyroid- simulating hormone)
 FSH (follice stimulating hormone)
 LH (luteinizing hormone)
Serum protein electrophoresis

Serum proteins are

separated into 6 groups:
Albumin

α1 - globulins
α2 - globulins
β1 - globulins
β2 - globulins
γ - globulins

Figure is found at http://www.sebia-usa.com/products/proteinBeta.html#

Common functions of plasma proteins
 Proteins are structural materials of animal body and
help in the growth of animal body
 buffer properties (maintenance of pH)
 build new tissues and maintain already present
tissues
 Some act as biochemical catalyst
 maintenance of osmotic pressure of blood
 Prevention of thrombosis (anticoagulant proteins)
 Defense against infection (antibodies, complement
proteins)
Albumin
 Functions include: HYPOALBUMINEMIA
 –Transport Common Causes
 –Osmotic pressure Decrease albumin synthesis:
regulation
a. Liver disease (specially
chronic diseases).
 •Synthesized in the liver. b. Malnutrition.
c. Alcoholism
 •Deficiency: in liver Increased albumin loss:
disease and kidney
disease. a. Renal disease (nephrotic
syndrome).
-Loss of albumin in urine
(proteinuria).
b. Extensive burns:
-Loss of albumin through
skin
 Non-protein Nitrogen Compounds
 Nitrogen containing compounds that are not proteins or
polypeptides
 The determination of non protein nitrogenous (NPN) substances
in the blood has traditionally been used to monitor renal
function.
 Useful clinical information is obtained from individual
components of NPN fraction
 Clinically signigicant NPN compounds are
urea,creatine,creatinine,uric acid etc.