PROTEIN/AMINO ACID

METABOLISM

Nitrogen balance
Protein content of adult body remains
remarkably constant
Protein constitutes 10-15% of diet
Equivalent amount of amino acids must be lost each
day

Amino acids pool

No storage facility for amino acids
Amino acids incorporated into functional proteins

Amino acids in blood and extracellular fluid

represent an amino acid pool
Amino acids move through this pool
Average 60 kg woman
10 kg protein
170 g free amino acids in pool

Fate of amino acids

If not required for protein synthesis amino groups removed
For most amino acids occurs primarily in liver
For Branced Chain Amino Acids (leucine, isoleucine, valine)
occurs primarily in skeletal muscle
amino groups transferred to alanine and taken to liver for disposal via
glucose-alanine cycle

Carbon skeletons used for:

Gluconeogenesis (in liver)
Oxidised in Krebs Cycle

Amino groups used for

Synthesis of nonprotein nitrogen compounds
disposed of via Urea Cycle

Figure Pool and Fate of Amino Acids in the Body

From: Summerlin LR (1981) Chemistry for the Life Sciences. New York: Random House p 563.

Metabolic relationship of amino acids

BODY PROTEINS
Proteosynthesis

NONPROTEIN
DERIVATIVES

GLYCOLYSIS
KREBS CYCLE
UREA

Conversion
(Carbon skeleton)

AMINO ACIDS

DIETARY
PROTEINS

GLUCOSE

Degradation

250 300
g/day

ACETYL CoA

NH3

CO2

Porphyrins
Purines
Pyrimidines
Neurotransmitters
Hormones
Komplex lipids
Aminosugars

KETONBODIES

Amino acid structure

The 20 common amino acids of proteins

Enzymes cleaving the peptide bond

Endopeptidases hydrolyse the peptide bond inside a
chain: pepsin, trypsin, chymotrypsin
Exopeptidases split the peptide bond at the end of a
protein molecule: aminopeptidase, carboxypeptidases

Dipeptidases
pepsin (pH 1.5 2.5) peptide bond derived from Tyr, Phe,
bonds between Leu and Glu
trypsin (pH 7.5 8.5) bonds between Lys a Arg

chymotrypsin (pH 7.5 8.5) bonds between Phe a Tyr

Essential amino acids in humans

Arginine*
Histidine*
Isoleucine
Leucine
Valine

Lysine
Methionine
Threonine
Phenylalanine
Tryptophan

*Required to some degree in young growing period and/or sometimes during illness.

Non-essential and nonessential

amino acids in humans
Can be formed from a-keto acids by transamination and
subsequent reactions.

Alanine
Asparagine
Aspartate
Glutamate
Glutamine

Glycine
Proline
Serine
Cysteine (from Met*)
Tyrosine (from Phe*)
* Essential amino acids

Amino acid metabolism

Metabolism of amino acids differs, but 3
common reactions:
Transamination

Deamination
Formation of urea

General reactions of amino acid catabolism

O
+ NH4+
1. Deamination

NH2
R

CH

COO-

2. Transamination

COO-

3. Urea Cycle

COO-

C
NH2

CH

COO-

oxidative
decarboxylation

NH3+
R

CH2

CO2

Transamination reactions
Amino group removed from one amino acid and
transferred to another
Catalysed by aminotransferase enzymes
Nearly all transaminations transfer amino group to aketoglutarate
Forms new ketoacid and glutamate (amino acid)

BCAAs transaminations in smooth muscles usually

result in formation of alanine (via glutamate)
Released from muscle
Allows amino groups from BCAAs to move from smooth
muscles to liver for disposal
BCAAs=Branced chain amino acids

Figure Diagram of transamination reactions of amino acids

From: Houston, ME. (2001) Biochemistry Primer for Exercise Science. Champaign: Human Kinetics. p151

Deamination reactions
Amino group (and H) removed
Forms ammonia (NH3)
Carbon skeleton left can be
Oxidised in Krebs Cycle
used for gluconeogenesis
converted to fatty acid

18 amino acids glucogenic/ketogenic

Leucine and lysine purely ketogenic

Utilization of carbon skleton from amino acids catabolism (deamination)

From: Houston, ME. (2001) Biochemistry Primer for Exercise Science. Champaign: Human Kinetics. p148

Urea cycle
Ammonia is toxic
Readily ionises to ammonium ion NH4+
NH4+ converted to urea in liver (urea cycle)
Urea contains 2 x NH2
One from NH4+
One from aspartate

Urea excreted in urine

Figure Urea Cycle

From: Stryer, LS (1988) Biochemistry (3rd Ed). New York: WH Freeman & Co. p500

The fate of the amino group during amino acid catabolism

Transamination reaction
The first step in the catabolism of most amino acids is
removal of a-amino groups by enzymes transaminases
or aminotransferases

All aminotransferases have the same prostethic group and

the same reaction mechanism.
The prostethic group is pyridoxal phosphate (PPL),
the coenzyme form of pyridoxine (vitamin B6)

Biosynthesis of amino acid:

transamination reactions
amino acid1 +a-keto acid2

amino acid2 +a-keto acid1

NH3+

O 2 CCH 2 CH 2 CHCO 2

Glutamate

O
R-CCO 2 -

Keto-acid

Pyridoxal phosphate (PLP)dependent aminotransferase

O
O 2 CCH 2 CH 2 CCO 2 -

a-Ketoglutarate

NH2
R-CHCO 2 -

Amino acid

Active metabolic form of vitamin B6

Mechanism of transamination reaction: PPL complex with enzyme accept

an amino group to form pyridoxamine phosphate, which can donate its amio
group to an a-keto acid.

All amino acids except threonine, lysine, and

proline can be transaminated
Transaminases are differ in their specificity for L-amino
acids.
The enzymes are named for the amino group donor.

Clinicaly important transaminases

Alanine-a-ketoglutarate transferase ALT
(also called glutamate-pyruvate transaminase GPT)
Aspartate-a-ketoglutarate transferase AST
(also called glutamate-oxalacetate transferase GOT)
Important in the diagnosis of heart and liver damage caused by heart
attack, drug toxicity, or infection.

ALT

Glucose-alanine cycle
Alanine plays a special role in
transporting amino groups to liver.
Ala is the carrier of ammonia and of the
carbon skeleton of pyruvate from muscle to
liver.
The ammonia is excreted and the pyruvate is
used to produce glucose, which is returned to
the muscle.

According to D. L. Nelson, M. M. Cox :LEHNINGER. PRINCIPLES OF BIOCHEMISTRY Fifth edition

Glutamate releases its amino group as

ammonia in the liver
The amino groups from many of the a-amino acids are collected in the
liver in the form of the amino group of L-glutamate molecules.

Glutamate undergoes oxidative deamination catalyzed by L-glutamate

dehydrogenase.
Enzyme is present in mitochondrial matrix.
It is the only enzyme that can use either NAD+ or NADP+ as the acceptor of reducing

equivalents.
Combine action of an aminotransferase and glutamate dehydrogenase referred to as

transdeamination.

Ammonia transport in the form of glutamine

Excess ammonia is added to
glutamate to form glutamine.

Glutamine
synthetase

Glutamine enters the liver and NH4+

is liberated in mitochondria by the
enzyme glutaminase.
Ammonia is remove by urea
synthesis.

Relationship between glutamate, glutamine

and a-ketoglutarate
NH3

NH3

glutamate

a-ketoglutarate
NH3

glutamine
NH3

A. Glutamate dehydrogenase

glutamate

NAD+

H2O

a-ketoglutarate

B. Glutamine synthetase (liver)

ATP

NH3

ADP

glutamine

C. Glutaminase (kidney)

glutamine

NH3

To urea cycle

From transamination
reactions

glutamate

H2O

glutamate

NH3

NADH

Oxidative deamination

A. Oxidative deamination
Amino acids

FMN

L-amino acid oxidase produces

+

H2O
L-amino acid oxidase

a-keto acids

+ FMNH2 +

NH3
O2
catalse

FMN
B. Nonoxidative deamination

H2O2

ammonia and a-keto acid directly,

using FMN as cofactor.
The reduced form of flavin must be
regenerated by O2 molecule.
This reaction produces H2O2
molecule which is decompensated by
catalase.
H2O

O2

Is possible only for hydroxy amino acids

serine

threonine
Serin-threonin dehydratase

pyruvate +

NH3

a-ketoglutate
+
NH3

Amino acid metabolism and central

metabolic pathways
20 amino acids are converted
to 7 products:
pyruvate
acetyl-CoA
acetoacetate
a-ketoglutarate
succynyl-CoA
oxalacetate
fumarate

Glucogenic Amino Acids

formed: a-ketoglutarate, pyruvate,
oxaloacetate, fumarate, or succinyl-CoA
Aspartate
Asparagine
Arginine
Phenylalanine
Tyrosine
Isoleucine

Methionine
Valine
Glutamine
Glutamate
Proline
Histidine

Alanine
Serine
Cysteine
Glycine
Threonine
Tryptophan

Ketogenic Amino Acids

formed acetyl CoA or acetoacetate

Lysine
Leucine

Both glucogenic and ketogenic amino

acids
formed: a-ketoglutarate, pyruvate,
oxaloacetate, fumarate, or succinyl-CoA in
addition to acetyl CoA or acetoacetate
Isoleucine
Threonine
Tryptophan
Phenylalanine
Tyrosine

The C3 family: alanine, serine, cysteine and

threonine are converted to pyruvate

Alanine

Serine

Cysteine

Threonine

Pyruvate

The C4 family: aspartate and asparagine are

converted into oxalacetate

Asparagine

Aspartic acid

Oxalacetate

The C5 family: several amino acids are converted into

a-ketoglutarate through glutamate

Glutamine

Proline

Arginine

Histidine

a-ketoglutarate

Interconversion of amino acids and intermediates of

carbohydrate metabolism and Krebs cycle

Metabolism of some selected

amino acids

Serine biosynthesis from glycolytic

intermediate 3-phosphoglycerate

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Glycine biosynthesis from serine

Reaction involves the transfer of the hydroxymethyl group from serine to the cofactor
tetrahydrofolate (THF), producing glycine and N5,N10-methylene-THF.
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Glycine oxidation to CO2

Glycine produced from serine or from the diet can also be oxidized by glycine
decarboxylase (also referred to as the glycine cleavage complex, GCC) to yield a
second equivalent of N5,N10-methylene-tetrahydrofolate as well as ammonia and
CO2.
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Cysteine and methionine are metabolically

related
The sulfur for cysteine synthesis comes from the essential amino acid
methionine.
SAM

Condensation of ATP and methionine

yield S-adenosylmethionine (SAM)

SAM serves as a precurosor for numerous methyl transfer reactions (e.g. the
conversion of norepinephrine to epinenephrine).

Cysteine synthesis
Conversion of homocysteine back to Met. N5methyl-THF is donor of methyl group.

*folate + vit B12

1.

Conversion of SAM to
homocysteine.

2.

Condensation of
homocysteine with serine to
cystathione.

3.

Cystathione is cleavaged to
cysteine.

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Homocystinuria
Genetic defects for both the synthase and the lyase.
Missing or impaired cystathionine synthase leads to homocystinuria.
High concentration of homocysteine and methionine in the urine.
Homocysteine is highly reactive molecule.
Disease is often associated with mental retardation, multisystemic
disorder of connective tissue, muscle, CNS, and cardiovascular
system.

Biosynthesis of Tyrosine from Phenylalanine

Phenylalanine hydroxylase is a mixed-function oxygenase: one atom of oxygen is

incorporated into water and the other into the hydroxyl of tyrosine. The reductant is the
tetrahydrofolate-related cofactor tetrahydrobiopterin, which is maintained in the reduced
state by the NADH-dependent enzyme dihydropteridine reductase

Phenylketonuria
Hyperphenylalaninemia - complete deficiency of phenylalanine
hydroxylase (plasma level of Phe raises from normal 0.5 to 2 mg/dL to
more than 20 mg/dL).
The mental retardation is caused by the accumulation of
phenylalanine, which becomes a major donor of amino groups in
aminotransferase activity and depletes neural tissue of -ketoglutarate.
Absence of -ketoglutarate in the brain shuts down the TCA cycle and
the associated production of aerobic energy, which is essential to
normal brain development.
Newborns are routinelly tested for blood concentration of Phe.
The diet with low-phenylalanine diet.

Catabolism of branched amino acids

valine

isoleucine

a-ketoglutarate

a-ketoisovalerate

leucine

glutamate (transamination)

a-keto-b-methylbutyrate

a-ketoisokaproate
NAD+

oxidative decarboxylation
Dehydrogenase of a-keto acids*

CO2

isobutyryl CoA

a-methylbutyryl CoA

NADH + H+

isovaleryl CoA

Dehydrogenation etc., similar to fatty acid b-oxidation

propionyl CoA

acetyl CoA
+
propionyl CoA

acetyl CoA
+
acetoacetate

Branched-chain aminoaciduria
Disease also called Maple Syrup Urine Disease (MSUD) (because
of the characteristic odor of the urine in affected individuals).

Deficiency in an enzyme, branched-chain -keto acid

dehydrogenase leads to an accumulation of three branchedchain amino acids and their corresponding branched-chain -keto
acids which are excreted in the urine.
There is only one dehydrogenase enzyme for all three amino
acids.
Mental retardation in these cases is extensive.

Histidine Metabolism:
Histamine Formation
H
N

+
NH3
CH2CHCO2 -

Histidine
decarboxylase

H
N

CH2CH2NH2

Histidine

CO2

Histamine

Histamine:
Synthesized in and released by mast cells
Mediator of allergic response: vasodilation,
bronchoconstriction

Tryptophan catabolism

Tryptophan has complex catabolic pathway:

1. the indol ring is ketogenic
2. the side chain forms the glucogenic products
Kynurenate and xanthurenate are excrete in the urine.

Enzymes which metabolised amino acides

containe vitamines as cofactors
THIAMINE B1 (thiamine diphosphate)
oxidative decarboxylation of a-ketoacids
RIBOFLAVIN B2 (flavin mononucleotide FMN, flavin adenine dinucleotide FAD)
oxidses of a-aminoacids

NIACIN B3 nicotinic acid (nikotinamide adenine dinucleotide NAD+

nikotinamide adenine dinukleotide phosphate NADP+)
dehydrogenases, reductase
PYRIDOXIN B6 (pyridoxalphosphate)
transamination reaction and decarboxylation
FOLIC ACID (tetrahydropholate)
Meny enzymes of amino acid metabolism

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