Protein and Amino Acid

Metabolism

Contessa S. Tac-an, M.D.

 OVERVIEW
 1ST Phase
-removal of alpha-amino groups forming
ammonia and alpha-ketoacids
 2nd Phase
-carbon skeletons converted to common
intermediates of energy-producing
metabolic pathways
Overall Metabolism of Protein Amino Acid
 Protein Turnover
 Rate of turnover varies.
 Chemical signals:
1. Ubiquitin
2. PEST sequence
Factors that affect the rate of utilization and
the proportion of the different amino acids

 Availability of other fuels

 Availability of exogenous amino acids
 The need of the organism for amino acids in
protein synthesis
 Nutrition dependence of the organism on
essential amino acids
 The need for specific amino acids as precursors
of other important biomolecules
 Central Role of the Liver in Amino Acid
Metabolism
Protein Synthesis of other Delivery to other
Synthesis N-CTG Compounds organs of a
balanced mixture
of amino acids

Catabolism of both Synthesis of

C-Chain and N of non-essential
amino acids amino acids
 Digestion of dietary proteins
1. Gastric secretion
a. HCl
b. pepsin
2. Pancreatic enzymes
> trypsin, chymotrypsin, elastase,
carboxypeptidases A & B.
3. Intestinal enzyme
> aminopeptidase
 Digestion of dietary protein
 Amino acid absorption
 Amino acid transport into cells
> ATP-requiring active transport systems
> e.g. COAL (cysteine, ornithine, arginine
and lysine) reabsorption in kidney tubules.
(Cystinuria – inherited deficiency in the
transport system in renal tubules.)
 Amino Acid Catabolism
A. Removal of the -NH2 group
1. Transamination
2. Deamination
2.a. Oxidative deamination
2.b. Non-oxidative deamination
B. Decarboxylation
C. Oxygenation
D. One-Carbon transfer
TRANSAMINATION
 Transamination
 Enzyme: Transaminase or aminotransferase
 General reaction

 Example

 Enzyme: Aspartate aminotransferase or Glutamate-oxaloacetate

transaminase
 Co-enzyme: Pyridoxal Phosphate (Vitamin B6)
Intermediate amino group
Carrier between the amino acid and the keto acid
 TRANSAMINATION
 Enzyme: Aminotransferases (transaminases)
 Except lysine and threonine
 Substrate specificity
> Alanine aminotransferase (ALT)
or glutamate pyruvate transaminase(GPT)
> Aspartate aminotransferase(AST)
or glutamate oxaloacetate transaminase(GOT)
 Aminotransferases
 Diagnostic value:
AST/SGOT & ALT/SGPT
> elevated in liver diseases and some
other non-hepatic diseases such as MI,
and muscle disorder.
 Oxidative Deamination

 Overall Reaction
Amino Acid Keto acid + NH3
 Enzymes involved
1. Glutamate dehydrogenase
COOH H 2O
COOH COOH
(CH2)2 + NAD+ (CH2)2 (CH2)2
CH-COOH NH=C-COOH O=C-COOH
NH2
-Iminoglutaric -Ketoglutarate
acid + NH4 + NADH2
Glutamate
 Oxidative Deamination (continued)

2. L-amino acid oxidase

L-a.a. + H2O + E-FMN -Keto acid + NH3 + E-FMNH2

3. D-amino acid oxidase

D-a.a. + H2O + E-FAD -Keto acid + NH3 + E-FADH2

R-CH-COOH + FP R-CH-COOH + FP. H2

NH2 Flavoprotein NH Reduced

enzyme enzyme
Amino acid Imino acid

R-CH-COOH + H2 O R-CH-COOH + NH3

NH O
Imino acid Keto acid

 Regeneration
FP. H2 + O2 FP + H2O2

2 H2O2 2 H2O + O2 Catalase

FP. H2 = E-FMNH2 or E-FADH2

 Non-Oxidative Deamination
 Enzymes involved
1. Amino acid dehydrases
 Requires B6PO4 as cofactor
 Hydroxy amino acids (serine, threonine,
homoserine)
-H20
OH-CH2-CH-COOH CH2=C-COOH CH3-C-COOH

NH2 NH2 NH
Imino acid
Serine Intermediate
±H2O
NH3 + CH3 -C-COOH

O
Pyruvic acid
 Non-Oxidative Deamination (continued)
2. Amino acid desulfhydrases
 Requires B6PO4 as cofactor
 Sulfur-containing amino acids (cysteine,
homocysteine)

-H2S
S-CH2-CH-COOH CH2=C-COOH CH3-C-COOH

NH2 NH2 NH
Cysteine Imino acid

NH3 + CH3 -C-COOH

O
Pyruvic acid
 Ways of Detoxifying Ammonia

1. Reversal of the glutamate dehydrogenase rxn

Glutamic acid -Ketoglutarate + NH3

2. Glutamine formation
Glutamine
Glutamic acid + NH3 Glutamine
Synthetase

3. Urea formation
4. Asparagine formation
Aspartic acid + NH3 Asparagine
 Urea Cycle

 Enzymes
1. Carbamoyl PO4
Synthetase
2. Ornithine
Transcarbamoylase
3. Argininosuccinate
synthetase
4. Argininosuccinase
5. Arginase
 UREA CYCLE
 Urea – major disposal form of amino
groups from amino acids.
- accounts for 90% of N-ctg.
components of urine.
- produced in the liver; excreted in
the urine.
 UREA CYCLE
 Sources of Nitrogen of Urea:
1. free NH3 2. Aspartate

 Glutamate – immediate precursor of NH3

& aspartate nitrogens
 CO2 – source of carbon & O2
 UREA CYCLE
 Reactions in Urea synthesis
Note: 1st 2 rxns occur in mitochondria,
then remaining cycle enzymes are
located in cytosol.
( Glutamate Dehydrogenase also occurs in
mitochondria to provide ammonia for
incorporation into carbamoyl phosphate.)
 UREA CYCLE
1. Formation of carbamoyl PO4
Enzyme: Carbamoyl phosphate
synthase
>requires N-Acetylglutamate
& cleavage of 2 ATP
2. Formation of Citrulline
Enzyme: Ornithine transcarbamoylase
(Ornithine & citrulline – basic a.a. used in
urea cycle.)
Citrulline transported to cytosol.
 UREA CYCLE
3. Synthesis of Argininosuccinate
Enzyme: Argininosuccinate synthase
Citrulline condenses w/ Aspartate (2nd
nitrogen source incorporated in urea).
> requires cleavage of ATP to AMP &
PPi.
4. Cleavage of Argininosuccinate
- yields arginine & fumarate
- Arginine : immediate precursor of urea
- Fumarate : hydrated to malate
 UREA CYCLE
5. Cleavage of Arginine to Ornithine & urea
Enzyme: Arginase (only found in liver)
Note: Only the liver can cleave Arginine &
synthesize urea.
6. Fate of urea  transported in the blood &
excreted in urine (or a small portion diffuses
from blood to intestine & cleaved to CO2 & NH3
by bacterial urease).
Renal failure  hyperammonemia
 Overall Stoichiometry of Urea Cycle

 Aspartate + NH3 + CO2 + 3 ATP 

Urea + fumarate + 2 ADP + AMP +
2 Pi + PPi + 3H2O
> 4 high energy phosphates consumed in
urea synthesis
> irreversible
> large, negative ΔG
 Regulation of Urea Cycle
 N-Acetylglutamate – activator for
carbamoyl PO4 synthetase 1; Rate limiting
step in urea cycle.

 Glutamate concentration in liver after each

protein meal.
 NH3 Metabolism
A. Sources of Ammonia
1. Amino acids
2. Glutamine
a. Renal enzyme glutaminase
- mostly excreted as NH4+ into urine;
maintenance of acid-base balance
b. Intestinal glutaminase
 Metabolism of Ammonia
3. Intestinal bacterial degradation of urea.
4. Amines
- hormones or neurotransmitter
- diet
- Enzyme Amine oxidase
5. Purines & Pyrimidines
 Transport of Ammonia in the
circulation
1. UREA
2. GLUTAMINE
- nontoxic storage & transport form of
ammonia
Glutamine formation occurs primarily in muscle & liver;
also important in nervous system where it’s the major
mechanism
for the removal of NH3 in brain.
- has high plasma concentrations due to its transport
functions.
- removed by the kidneys & deaminated by glutaminase
 Metabolism of Ammonia
 Hyperammonemia/ammonia intoxication
S/Sx: tremors, slurring of speech, blurring
of vision
> coma & death
1. Acquired hyperammonemia
e.g. liver cirrhosis (Alcoholism, hepatitis,
biliary obstruction)
 collateral circulation (portal blood
shunted to systemic circulation)
 Metabolism of Ammonia
2. Hereditary hyperammonemia
> genetic deficiencies of 5 enzymes of
urea cycle (1 in 30,000 live births)
> mental retardation
 Mechanism of NH3 Toxicity
 Shift in equilibrium of glutamate DH rxn
toward glutamate formation
 Alpha-ketoglutarate + NADPH + H+ + NH3
Glutamate + NADP+
 Depletes Alpha-ketoglutarate  lowers
ATP production
 (Brain is vulnerable because it depends on
CAC for energy production.)
Catabolism of Carbon skeletons of
amino acids
Catabolic Disposition (Fates) of Carbon
Chains of Amino Acids
 Metabolic Fates of the Keto Acids

1. Synthetic pathway
-Ketoacid + NH3 -amino acid

2. Glucogenic pathway
3. Ketogenic pathway
4. Miscellaneous pathways

 Ketogenic amino acid - Leucine

Metabolic Fates of the Keto Acids
 Both glycogenic and  Glycogenic amino
ketogenic acids

 Ile  Ala Val

 Phe  Arg Met
 Tyr  Asp Gly
 Lys  Asn His
 Trp  Cys Ser
 Glu Thr
 Gln Pro
AAs that form oxaloacetate
AAs that form Alpha-ketoglutarate
1. Glutamine  glutamate + NH3
Glutamate  alpha-ketoglutarate
2. Proline --- Δ1-pyrroline 5 –
carboxylate - glutamate  alpha-KG
3. Arginine  ornithine  glutamate-γ-
semialdehyde  alpha KG
4. Histidine  N-formiminoglutamate 
glutamate + formiminotetrahydrofolate
AAs that form pyruvate
AAs that form pyruvate
 AAs that form pyruvate
1. Alanine
2. Serine
3. Glycine
4. Cystine + NADH+H+  cysteine
(undergoes desulfuration to yield
pyruvate)
5. Threonine  pyruvate or alpha-
ketobutyrate  succinyl CoA
 Pathways of Degradation of Glycine

1. Major Route – Glycine synthase

Glycine + FH4 + NAD N5N10 –Methylene FH4
+ CO2 + NH3 + NADH + H+

2. Conversion to serine by serine hydroxymethyl

transferase
3. Oxidative deamination by glycine oxidase to
yield glyoxylic acid
H 2O Glycine H2O2
oxidase
Glycine Glyoxylic acid

O2 NH3
 Metabolic Pathways for Glycine

 Heme synthesis
 Synthesis of purines  forms positions 4, 5, 7 of
the purine ring
 Constituent of glutathione
 Conjugates with cholic acid to form glycocholic
acid
 Conjugates with benzoic acid to form hippuric
acid
 Synthesis of creatine
 Two Principal Catabolic Pathways of
Cysteine
1. Direct oxidative pathway-
 Cysteine sulfinate pathway

Cysteine Cysteine sulfinate pyruvate

2. Transamination pathway-
 3-Mercaptopyruvate pathway

Cysteine 3-Mercaptopyruvate pyruvate

 Inborn Errors

1. Cystinuria (Cystine-Lysinuria)
 Considered to be due to a renal transport
defect affecting renal reabsorptive
mechanisms for 4 amino acids – cystine,
lysine, arginine and ornithine
 Manifested by increased urinary excretion of
cystine, lysine, arginine and ornithine
 Cystine is insoluble  may precipitate in
kidney tubules and form cystine calculi
 Inborn Errors (continued)

2. Cystinosis (Cystine Storage Disease)

 Primary defect: Impaired lysosomal function
 Signs and symptoms
 Deposition of cystine crystals in many
tissues and organs particularly the
reticuloendothelial system
 Impaired renal function leading to acute
renal failure
 Pathways for Threonine Degradation

1. Conversion to -Ketobutyric acid by threonine

dehydratase
-Ketobutyric acid propionic acid glucose

2. Cleavage to glycine and acetaldehyde by

threonine aldolase
 Pathways for Threonine Degradation
(continued)
3. Dehydrogenation and decarboxylation to yield
aminoacetone

Aminoacetone + O2 2-Ketopropanol Pyruvate

Aldehyde DH
Acetaldehyde Acetyl CoA

NAD+ CoA NADH

AAs that form Fumarate
 Inherited enzyme deficiency
 PKU
 Alkaptonuria
 Albinism
 Metabolic Pathway of Phe/Tyr

 Enzymes
1. Phenylalanine
monooxygenase
or phenylalanine
oxidase
or phenylalanine
hydroxylase
2. Homogentisate
1,2-dioxygenase
3. Tyrosinase
 AAs converted to Succinyl CoA
1. Methionine
2. Valine & Ile
3. Threonine  alpha-ketobutyrate 
propionyl CoA  succinyl CoA or to
pyruvate
AAs converted to succinyl CoA
 Methionine

 Inborn Errors
1. Homocystinuria
 Deficiency to
cystathionine
synthetase
2. Cystathioninuria
 Deficiency to
cystathionase
 -KG
Transamination
Glu
-Ketoisovaleric acid
CoA, NAD+
-Ketoisovaleric acid
dehyrogenase NADH, CO2
Isobutyryl CoA
Acyl CoA DH
Methylacrylyl CoA

Methylmalonic acid semialdehyde

Propionyl CoA

Succinyl CoA
 HMG-CoA
 obligatory intermediate in cholesterol
biosynthesis and ketogenesis

 Maple Syrup Urine Disease

 Deficiency of -keto acid dehydrogenase
 -KG
Transamination
Glu
-Keto-ß-methylvaleric acid
CoA, NAD+
-Ketoisovaleric acid
dehyrogenase NADH, CO2
-Methylbutyryl CoA
Acyl CoA DH
Tiglyl CoA

-Methylacetoacetyl CoA

Propionyl CoA

Methylmalonyl CoA

Succinyl CoA
 AAs converted to Acetyl CoA or
Acetoacetyl CoA
1. Leucine
2. Ile
3. Lys  Alpha-aminoadipate-semialdehyde
Acetoacetyl CoA
4. Tryptophan
 Leucine -KG
Transaminase
Glu
-Ketoisocaproic acid
CoA, NAD+
-Ketoisocaproic
dehyrogenase NADH, CO2
Isovaleryl CoA
Acyl CoA DH

ß-Methylcrotonyl CoA

ß-Hydroxy-ß-methyl
Glutaryl CoA (HMG-CoA)

Acetyl CoA Acetoacetic acid

 L-amino acid Saccharopine
oxidase DH
L-Ketoaminocaproic Saccharopine
acid
Saccharopine H2O, NAD+
Pipecolic acid DH
NADH,
L-Glutamate
L--Aminoadipic acid semialdehyde

-Aminoadipic acid

-Ketoadipic acid

Glutaryl CoA

Crotonyl CoA

Acetoacetyl CoA
Glucose – Alanine Cycle
 Functions of Glucose – Alanine Cycle

1. To carry amino groups from skeletal muscle to

the liver to be converted to urea.
2. To provide the working muscle with blood
glucose made by the liver from the carbon
backbone of alanine.
3. Smooths out fluctuations in the blood glucose
level in the periods between meals.
 Nitrogen balance

Energy, heme, purines, etc.

Dietary Amino acid Excreted as urea,

protein pool NH4+

Tissue
Protein
 Nitrogen Balance

 Negative nitrogen balance

 Due to metabolic stress
 Due to lack of an essential amino acid
 Inadequate dietary protein
 Starvation
 Positive nitrogen balance
 Growth
 Pregnancy
 Convalescence