PROTEIN METABOLISM

PRASHANT REGMI
Introduction

• Unlike fats and carbohydrates, amino acids are not stored by

the body.
• No protein in body exists whose sole function is to maintain a
supply of amino acids for future use.
• Therefore, amino acids must be obtained from the diet,
synthesized de novo, or produced from normal protein
degradation.
• Any amino acids in excess of the biosynthetic needs of the cell
are rapidly degraded.
• The first phase of amino acid catabolism involves the removal
of the α-amino groups, forming ammonia and the
corresponding α-keto acid.
• A portion of the free ammonia is excreted in the urine, but
most is used in the synthesis of urea.
• In the second phase of amino acid catabolism, the carbon
skeletons of the α-keto acids are converted to common
intermediates of energy producing, metabolic pathways.
Amino acid pool

• Amino acids are continuously

obtained either from:

– Dietary proteins
– Degradation of tissue proteins
– Synthesis of non-essential amino
acids
Amino acid pool

• Amino acids are continuously

utilized for biosynthetic
reactions:
– Formation of tissue proteins
– Synthesis of porphyrins, creatine,
neurotransmitters & other
nitrogen containing compounds
– Used to form glucose/ glycogen
– Used to form ketone bodies, fatty
acids or steroid
– Amino acids not used in
biosynthetic reactions are burned
as fuel.
Amino acid pool

• Amino acids formed in body and consumed in body constitute

amino acid pool.
• Though, amino acids are not stored in body, amino acid pool is
in dynamic state.
• About 400 g from tissue and about 100 g from dietary protein
enter into amino acid pool per day which is equivalent to about
100 g amino acids.
• About total (400 g/day) of tissue proteins are regenerated under
normal condition, 30 g/day go for synthesis of specialized
products.
• Amount burned as fuel or converted to storage molecules varies.
Amino acid pool

• Digestion of dietary proteins

• Protein turnover
– Cellular proteins
– Extra-cellular proteins
Digestion of dietary proteins

• Most of the nitrogen in the diet is consumed in the forms of

protein ( about 70-100 g/day).
• Proteins are too large to be absorbed by the intestine.
(Exception: newborns can take up maternal antibodies in
breast milk.)

• So, these proteins must be hydrolyzed to yield their constituent

amino acids, which can be absorbed.
Protein turnover

• Most proteins in the body are constantly being synthesized and

then degraded, which permit the removal of abnormal or
unneeded proteins.
• Cells have mechanisms for detecting and removing damaged
proteins.
• A significant proportion of newly synthesized protein
molecules are defective because of errors in translation.
• Even proteins that are normal when first synthesized may
undergo oxidative damage or be altered in other ways with the
passage of time.
Protein turnover

• There are two major enzyme systems responsible for degrading

damaged or unneeded proteins:
– The energy-dependent Ubiquitin-proteasome mechanism
This system mainly degrade endogenous proteins, that is ,
proteins that were synthesized within cell.

– Non-energy dependent degradative enzymes of the lysosomes

This system primarily degrade extra cellular proteins, such as
plasma proteins that are taken into the cell by endocytosis,
and cell-surface membrane proteins that are used in receptor-
mediated endocytosis.
• The ubiquitin is
recycled and the
peptide products are
further degraded by
other cellular
proteases to yield
individual amino
acids.
Lysosomal degradation of proteins

• Extracellular proteins are degraded by enzyme systems of

lysosome.
• Intra-lysosomal proteases hydrolyze internal peptide bonds.
• The resulting peptides are then degraded to amino acids by
endopeptidases that cleave internal bonds and by
aminopeptidases and carboxypeptidases that remove amino
acids sequentially from the amino and carboxyl terminals,
respectively.
Catabolism of amino acids

• The presence of the α-amino group keeps amino acids

safely locked away from oxidative breakdown.
• Removing the α-amino group is essential for producing
energy from any amino acid, and is an obligatory step in the
catabolism of all amino acids.
• Once removed, this nitrogen can be incorporated into other
compounds or excreted, with the carbon skeleton being
metabolized.
Transamination

• The purpose of transamination is

funneling of amino groups of
amino acids to glutamate.
• The first step in the catabolism of
most amino acids is the transfer of
their α-amino group to α-
ketoglutarate.
• The products are an α-keto acid
(derived from original amino
acid), and glutamate.
• α-keto glutarate plays a unique role
in amino acid metabolism by
accepting the amino groups from
other amino acids, thus becoming
glutamate.
• Glutamate produced by
transamination can be oxidatively
deaminated, or used as an amino
group donor in the synthesis of
non-essential amino acids.
• This transfer of amino groups from
one carbon skeleton to another is
catalyzed by a family of enzymes
called aminotransferases.
• All amino acids, with the exception of
lysine and threonine, participate in
transamination at some point in their
catabolism.
• There is no net deamination in these
reactions because the α-ketoglutarate
becomes aminated as the α-amino acid
is deaminated.
• The effect of transamination reactions is to collect the
amino groups from different amino acids in the form of L-
glutamate.
• Cells contain different types of aminotransferases, many are
specific for α-ketoglutarate as the amino group acceptor but
differ in their specificity for the L-amino acids.
Two most important amino transferases are-

1. ALT:
– Formerly called glutamate-pyruvate transaminase (GPT)

2. AST:
– Formerly called glutamate-oxaloacetate transaminase (GOT)
– AST transfers amino groups from glutamate to oxaloacetate,
forming aspartate, which is used as a source of nitrogen in the
urea cycle.
Diagnostic value of aminotransferases

• Aminotransferases are normally intracellular enzymes, with

the low levels found in the plasma representing the release
of cellular contents during normal cell turnover.
• The presence of elevated plasma levels of aminotransferases
indicates damage to cells rich in these enzymes.
• Two aminotransferases- ALT & AST- are of particularly
diagnostic value.
Liver disease

• Plasma ALT & AST are elevated in nearly all lever

diseases.
• ALT is more specific for liver disease.
• AST is more sensitive because the liver contains larger
amounts of AST.

• ALT/AST = >1, in most liver diseases.

• ALT/AST = <1 in necrotic diseases like cirrhosis and

alcoholic liver diseases.
Deamination

• The amino groups from many of the α-amino acids are

collected in the liver in the form of the amino group of
glutamine molecules.
• These amino groups must next be removed from glutamate
to prepare them for excretion.
• In hepatocytes, glutamate is transported from the cytosol
into mitochondria, where it undergoes oxidative
deamination catalyzed by L-glutamate dehydrogenase (MW
330 kD).
• It is the only enzyme that can use either NAD+ or NADP+
as the acceptor or reducing equivalents.
D-amino acid Oxidase

• D-amino acid are found in plants & in the cell walls of

microorganisms, but are not used in the synthesis of
mammalian proteins.
• D-amino acids are, however, present in the diet, and are
efficiently metabolized by the liver.
• D-amino acid oxidase is an FAD-dependent enzyme that
catalyzes the oxidative deamination of these amino acid
isomers.
• The resulting α-ketoacids can enter the general pathways of
amino acid metabolism & be reaminated to L-isomers, or
catabolized for energy.
Ammonia metabolism

• Ammonia is produced by all tissues during metabolism of a

variety of compounds, and is to be transported to liver for
the formation of urea.
• However, the level of ammonia in the blood must be kept
very low, because even slightly elevated concentration are
toxic to the CNS.
• Therefore, there must be a metabolic mechanism by which
nitrogen is moved from peripheral tissues to liver for
ultimate disposal as urea, while at the same time low levels
of circulating ammonia must be maintained.
Sources of ammonia
1. From amino acids
2. From glutamine
3. From bacterial action in intestine
4. From amines
5. From purines and pyrimidines
Transport of ammonia to Liver

• In most animals much of the free

ammonia is converted to a nontoxic
compound before export from the
extra-hepatic tissues in the blood
and transport to liver or kidney.
• The free ammonia produced in
tissues is combined with glutamate
to yield glutamate by the action of
glutamine synthetase.
• This reaction requires ATP &
occurs in two steps.
Glutamine

• Glutamine is a non-toxic transport form of ammonia; it is

normally present in blood in much higher concentration
than other amino acids.
• Glutamate also serves as a source of amino groups in a
variety of biosynthetic reactions.
• Glutamine in excess of that required for biosynthetic
reaction is transported in the blood to the intestine , liver,
and kidney for processing.
• In these tissues, the amide nitrogen is released as
ammonium ion in the mitochondria, where the enzyme
glutaminase converts glutamine to glutamate & NH4+.
• The NH4+ from intestine &
kidney is transported in the
blood to the liver.
• In liver, the ammonia from all
sources is disposed off by urea
synthesis.
Glucose-Alanine cycle

• In muscle and certain

other tissues that
degrade amino acids
for fuel, amino groups
are collected in the
form of glutamate by
transamination.
• The capacity of the hepatic urea cycle exceeds the normal
rates of ammonia generation.
• However, when the liver function is compromised or
there is any genetic defect, the blood ammonia level can
rise.
• Such hyperammonemia is a medical emergency, because
ammonia has a direct neurotoxic effect on the CNS.
Overall metabolism of ammonia
Urea Cycle

• If not reused for the synthesis of new amino acids or other

nitrogenous products, amino groups are channeled into a
single excretory end products.

– Ammonotelic Ammonia

– Ureotelic Urea

– Uricotelic Uric acid

Krebs bicycle