Amino Acids: Disposal of Nitrogen

◼ Amino acids are not stored in the body

◼ AA obtained from:

1. Diet
2. de novo synthesis
3. protein degradation
▪ Nitrogen leaves the body as
▪ Urea, ammonia & product from AA
catabolism
Amino Acid Pool
◼ Protein turnover:
balance between degradation and synthesis makes
total body protein constant
▪ Protein degradation:
two major enzyme degradation systems for damaged
body proteins:
▪ Energy-dependent ubiquitin-proteasome mechanism
for endogenous proteins
▪ Lysosomal non energy-dependent degradative
enzymes for extracellular protein e.g plasma protein,
cell surface protein
Digestion of Protein
◼ Begins in the stomach (HCL +pepsinogen)
◼ Pancreatic enzymes (proteases) activated by
cholecystokinin & secretin hormones ( polypeptide)
◼ Digestion of oligopeptides by small intestine enzymes
(exopeptidase from N-terminal also called
aminopeptidase)
◼ Absorption of free AA & dipeptides from small
intestine via portal circulation to go to liver
◼ Abnormalities in protein digestions:
 pancreatic enzymes→ appearance of undigested
proteins and fat ( steatorrhea) in feces.
Digestion of Protein
◼ Proteolytic enzymes: responsible for protein
degradation and produced by three different
organs (Stomach, pancreas, small intestine)
◼ Endopeptidase: Enzymes acting on the middle
of polypeptide chains.
◼ Exopeptidase: Enzymes acting on the
periphery of polypeptide chains.
◼ (1) In stomach: (HCL +pepsinogen)
 Digestion of Protein

(2) In the intestine :Proteolytic enzymes

endopeptidase exopeptidase Proteases

1) Trypsine 1)Procarboxypeptidase 1)Aminopeptidase
2) Chemotrypsine 2)Carboxypeptidase A 2)Dipeptidase
3) Elastase 3)Carboxypeptidase B 3) Tripeptidase
Transport of Amino Acids
◼ (1) Active transport
◼ with 7 transport systems according to tissue types.
Energy required system involves the uptake by cell of
a sodium ion and AA by the same carrier protein
◼ (2) Glutamyle cycle:
◼ AA reacts with Glutathion ( Glutamate, Cysteine,
Glycine) at the surface of cell membrane attached to
glutamate and released into the cytoplasm .
◼ Then the glutamyle moiety is used in the re-synthesis
of glutathion.
Cystinuria
◼ Abnormal AA transport system:
Cystinuria: defect in transport system
for cyn, ornithine, arg & lys (COAL) in
kidney→ precipitation of cyn in
kidney→ kidney stone of cyn type (
genetic disease)
Cystinuria
Removal of Nitrogen from AA
◼ Transamination: transfer of NH2 from one carbon
skeleton to another (funneling of NH 2 to glu)
◼ Enzymes: aminotransferases or transaminases
◼ Found in cytosol of liver, kidney intestine & muscle
◼ All AA except Lys & Thr participate in transamination.
Lys & Thr lose α-NH2 by deamination
◼ Examples: Alanine aminotransferase (ALT) formally
known as GPT
◼ Aspartate aminotransferase (AST) formally known as
GOT
Transamination Requires
Pyridoxal phosphate (B6)
Diagnostic Value of Aminotransferase

◼ Cell damage→ AST & ALT in blood

◼ Liver diseases: ALT is more specific
than AST but AST is more sensitive
because liver has more AST
◼ Myocardial infarction
◼ Muscle disorder
Oxidative Deamination of AA
◼ Liberation of NH2 as NH3 in liver &
kidney
◼ Enzyme : Glutamate DH

◼ Coenzyme: NAD or NADP

▪ Regulation :ATP & GTP allosteric

inhibitors
ADP & GDP allosteric activators
Oxidative Deamination of AA
Combined actions of aminotransferase & Glu
Dehydrogenase

◼ After meal disposal of AA proceed (A)

▪ Synthesis of AA proceed in case of
AA need (B)
Combined actions of aminotransferase &
Glu Dehydrogenase
D-Amino Acid Oxidase
◼ D-AA is found in diet from plant, cell
membrane of microorganism & body
must changed into
α L-AA by FAD-
dependent oxidase to be used for
synthesis of mammalian protein
◼ D-AA +O2→α-ketoacid + H2O2 + NH3
◼ α-ketoacid can either catabolized for
energy or converted to L-AA
Transport of NH3 in different tissues to
liver
Urea Cycle (NH2)2C=O
◼ Major disposal of amino groups of AA
◼ Urea is produced by liver→ blood→ kidney to
be excreted
◼ Need NH3 +CO2+Asp+ATP→ urea+ fumarate
◼ NH3 is from Glu (oxidative deamination)
◼ C & O is from CO2
◼ One N of urea is from NH3 and the other from
Asp ( OAA →Asp by AST)
y
Reactions of Urea Cycle
◼ Two reactions occur in mitochondria and the
others occur in cytosol
◼ Rate limiting step is carbamoyl phosphate
synthetase I & allosterically activated by N-
acetyl glutamate
◼ Enzyme is increased after protein rich diet to
provide Glu & increases urea synthesis
◼ four ATP are consumed( 2ATP→2ADP & ATP
→AMP +PPi) & urea synthesis is irreversible
N.B on Urea Cycle
◼ Fumarate produced by urea cycle→
malate shuttle →to mitochondria→ TCA
◼ Or fumarate in cytosol→ malate→

OAA→Asp
▪ Kidneys can synthesize Arg from
citrulline & Asp but liver is the only
organ to cleaves Arg into urea &
ornithine by arginase
Fate of Urea
◼ Urea in liver diffuses →blood →kidney→
urine to be excreted
◼ Urea in blood→intestine→CO2 + NH3 by
bacterial urease → feces or reabsorbed into
blood
◼ In kidney failure, plasma & gut urea levels
increase →NH3 → hyperammonemia
◼ Neomycin administration NH3 due to 
intestinal bacteria inhibiting urease
Flow of N from AA to Urea

▪ Amino groups for urea

synthesis are collected
in the form of ammonia
and aspartate
Metabolism of Ammonia
◼ Produced by all tissues during
metabolism
◼ Disposed primarily by urea cycle
◼ Blood NH3 must be low because of its
toxicity to CNS
◼ NH3 from peripheral tissues→ liver→
for urea cycle
Sources of Ammonia
◼ From AA by transamination & oxidative
deamination
◼ From Gln in kidney & intestine by glutaminase
→ NH3 → NH4+ to maintain acid-base balance
◼ From bacterial urease in intestine urea→ NH3
→ portal vein →liver
◼ From amines of diets & monoamines from
hormones & neurotransmitters by amine
oxidase
◼ From purines & pyrimidines catabolism
Transport of Ammonia
◼ Muscle releases AA nitrogen in the form
of Gln or Ala rather than in form of NH3
◼ Liver formation of urea → kidney
◼ Liver, muscle & NS (major mechanism)
can dispose NH3 as Gln by glutamine
synthetase using Glu
◼ Therefore, plasma Gln is found in
higher concentration than other AA
Glutamine/Glutamate
Metabolism of Ammonia
Hyperammonemia
◼ Normal serum level of NH3 5-50 μmol/L
◼ Defects in urea cycle enzymes or liver
diseases → ↑ NH3 in blood above 1000
μmol/L → hyperammonemia
◼ Symptoms: neurotoxic, tremors, slurring
speech, vomiting, cerebral edema & coma→
death
◼ Types: 1. acquired, due to liver diseases
2. hereditary, defect in urea enzymes
Hereditary
Hyperammonemia
◼ Mechanism of toxicity:
shift to synthesize more
glu from α-ketoglutarate
depleting it from TCA→
 Oxidation & ATP
▪ Brain is most affected due
To its dependency on TCA
For energy production