UREA CYCLE

PREPARED BY: CST, RMT 2020

Review
1. Glutamate production via transamination.
→α-ketoglutarate + α-amino acid → GLUTAMATE + α-keto acid
→α-ketoglutarate + alanine → GLUTAMATE + pyruvate
2. Glutamate to Aspartate via 2nd transamination
3. Ammonium ion production via oxidative deamination
©ResearchGate
UREA CYCLE
• The net effect of transamination and deamination reactions is production
of ammonium ions and aspartate molecules.
• Ammonium ions (1st reaction in Urea Cycle)→ entered in the urea cycle
indirectly as carbamoyl phosphate.
• Aspartate (2nd reaction in Urea Cycle)→ entered in the urea cycle directly.
*Carbamoyl phosphate & aspartate are “fuels” for the urea cycle.
UREA CYCLE
• The UREA CYCLE is a cyclic biochemical
pathway in which urea is produced, for
excretion, using ammonium ions and aspartate
molecules as nitrogen sources.
• Urea cycle happens in the liver→ Urea →blood
→kidney → urine

©Toppr.com
UREA
•
•
•
•
•

Pure state→ white, melting point of 133C

Soluble in water (1g/1ml)
Odorless
Colorless
Salty taste

• Adult excretes 30g of urea daily in urine/ varies with the protein content
of the diet.
INTERMEDIATES OF UREA CYCLE
• Arginine (standard)- *the most nitrogenrich of the standard AA
• Citrulline (nonstandard)
• Ornithine (nonstandard)

ARGININE

© American Chemical Society

CITRULLINE

© AdooQ Bioscience

*Nonstandard amino acids are not found

in protein.
ORNITHINE

© MedChemExpress
CARBAMOYL PHOSPHATE
Reactants:
• Ammonium ion
• CO2 in the form of bicarbonate(from
Krebs cycle)
• Water
• 2 ATP molecules
• Enzyme: Carbamoyl phosphate
synthetase I

Happens mitochondrial matrix

©Chegg
CARBAMOYL PHOSPHATE
• The formation of carbamoyl phosphate is stimulate by N-acetylglutamate
(required allosteric activator).
Glutamate + Acetyl-CoA=> N-acetylglutamate
• Enzyme: N-Acetylglutamate synthetase
• Positive regulator: Arginine
• Increased Amino acids→ increased arginine→ increase N-acetylglutamate
synthetase activity→ increased urea cycle action→ increased urea
production.
©ResearchGate
© Research Gate
STEPS OF THE UREA CYCLE
•
•
•
•

Step 1. Carbamoyl group transfer

Step 2. Citrulline-aspartate condensation
Step 3. Argininosuccinate cleavage
Step 4. Urea from arginine hydrolysis

*Step 1: Mitochondrial matrix

*Steps 2-4: Cytosol
Step 1. Carbamoyl group transfer
• Carbamoyl phosphate transfers its carbamoyl group (amide
group) to ornithine to form citrulline.
• Products: Pi & citrulline
• Enzyme: Ornithine transcarbamoylase

©Slideplayer
Step 1. Carbamoyl group transfer
• The breaking of high-energy phosphate
bond in carbamoyl phosphate drives the
transfer process.
• The 1st of the 2 nitrogen atoms and
carbon atom needed to form urea have
been introduced into the cycle.
Step 2. Citrulline-aspartate condensation
• Condensation between citrulline and aspartate occurs.
• Products: argininosuccinate, AMP, Ppi
• Enzymes: argininosuccinate synthetase

©Slideplayer
Step 2. Citrulline-aspartate condensation
• The 2nd nitrogen of the 2 nitrogen atoms to form urea has been
introduced into the cycle.
Glutamate dehydrogenase

NH4+

GLUTAMATE

Carbamoyl
phosphate

Oxaloacetate

Aspartate
Aminotransferase

Argininosuccinate
Step 3. Argininosuccinate cleavage
• Argininosuccinate is converted to arginine (standard amino acid) and fumarate
(intermediate of Krebs cycle)
• Enzymes: argininosuccinate lyase

©Slideplayer
Step 4. Urea from arginine hydrolysis
• Arginine is hydrolyzed, producing urea and regenerates
ornithine.
• Enzyme: Arginase

©Slideplayer
Step 4. Urea from arginine hydrolysis
• The oxygen atom in the urea comes from the water involved in
the hydrolysis.
• The ornithine is transported back into the mitochondria.

©Slideplayer
Urea Cycle Net Reaction
NH4+ + CO2 + 3 ATP + 2 H2O + aspartate =>
*urea + 2 ADP + AMP + Ppi + 2 Pi + fumarate
*Products
4 ATPS are consumed
• 2 ATP molecules are consumed in the formation of carbamoyl
phosphate.
• Equivalent of 2 ATP molecules is consumed in step 2 where 1 ATP is
hydrolyzed to AMP and PPi and the PPi is then further hydrolyzed to 2
Pi.
N1

N3
N1
N2
N4
©Bartleby.com

N4
REACTANTS

ENZYME

PRODUCT

Glutamine

Glutaminase

Glutamate, NH3

Glutamate

N-Acetylglutamate
synthetase

N-acetylglutamate

N-acetylglutamine
(activator), NH3, HCO3,
2ATP, H2O

Carbamoyl phosphate
synthetase I

Carbamoyl phosphate

Carbamoyl phosphate,
Ornithine

Ornithine transcarbamylase

Citrulline

Citrulline

Argininosuccinate

Argininosuccinate

Argininosuccinate
synthetase
Argininosuccinate lyase

Arginine

Arginase

Ornithine

Arginine
UREA CYCLE DISORDER
• Ammonia NH3 (neurotoxin)
• Normally, free ammonia is fixed into either α-ketoglutarate or
glutamine.
• Increased glutamine→ increased NH3 in the blood
(hyperammonemia)
• Increased NH3 can accumulate in the brain, causing confusion,
coma, or even death.
©Slideshare
Causes of Increased Ammonia
• Genetics (deficiency in the liver enzymes)
• Acquired hepatic causes (liver cirrhosis)
©Slideshare
Urea Cycle (ND) Retrieved from
http://education.med.nyu.edu/mbm/aminoAcids/ureaCycl
e.shtml, on October 18, 2020
Stoker, H.S.(2015). Biochemistry, 3rd ed. Quezon City. C&E
Publishing