AMMONIA METABOLISM

UREA CYCLE

Compiled by:-
PRATEEK CHOPRA
BT/BIO/05/310022
AMITY INSTITUTE OF BIOTECHNOLOGY
NOIDA
 OBJECTIVES

1. Define protein balance, nitrogen balance and essential amino acid.

2. Describe the transaminase, and glutamate dehydrogenase reactions

and discuss their roles in the removal of nitrogen waste in the body.

3. Identify the direct sources of nitrogen for the urea cycle.

4. Define hyperammonemia and discuss why a defect in either carbamoyl

phosphate synthetase I or ornithine transcarbamoylase leads to
hyperammonemia

5. Distinguish between ketogenic and gluconeogenic (glycogenic) amino

acids.

6. Describe the phenylalanine hydroxylase reaction and explain its

relationship to phenylketonuria;
PHYSIOLOGICAL PREMISE

Have you ever carefully read a packet of EqualTM? If so,

you may have noticed a warning to phenylketonurics. The
chemical sweetener in equal is a dipeptide containing
phenylalanine and aspartate. Some individuals are born with
one of the more common amino acid disorders,
phenylketonuria. They are unable to metabolize
phenylalanine to tyrosine. Consequently vast amounts of
phenylalanine will accumulate in the blood if too much of this
amino acid is consumed in the diet. Constant excess of
phenylalanine in the blood can cause severe mental
retardation. Hence this is one of several diseases tested for
in newborns in all states.
 Table 1- The essential and non-essential amino acids

Essential Nonessential
Argininea Methionineb Alanine Glutamine
Histidine Phenylalaninec Aspartate Glycine
Isoleucine Threonine Asparagine Proline
Leucine Tryptophan Cysteine Serine
Lysine Valine Glutamate Tyrosine

a
Arg is synthesized in the urea cycle, but the rate is too slow to meet
the needs of growth in children

b
Met is required to produce cysteine if the latter is not supplied
adequately by the diet.

c
Phe is needed in larger amounts to form tyr if the latter is not supplied
by the diet.
 BODY PROTEIN

Proteolysis Protein synthesis

De novo
Carbon compounds synthesis Catabolism
+ nitrogen Amino Acid Pool Urea + CO2

Dietary amino Biosynthesis of

acids nitrogen compounds
Porphyrins, creatine, carnitine,
hormones, nucleotides

Amino acid sources

Fates of amino acids

Figure 1. Sources and fates of amino acids

 PROTEIN BALANCE
• positive: synthesis > degradation (e.g., growth, body building)
• negative: synthesis < degradation (e.g., starvation, trauma, cancer cachexia)

BODY PROTEIN

Proteolysis Protein synthesis

Amino Acid Pool

 NH2 O

HOOC-CH-R HOOC-C-CH2CH2COOH
α-Amino acid α-Ketoglutarate

Cofactor = pyridoxal phosphate

α-Keto acid Glutamate

O NH2
HOOC-C-R HOOC-CH-CH2CH2COOH

Figure 2. Depiction of a general transamination

(aminotransferase) reaction. The α-amino acid other than
glutamate can be a wide variety
 Aspartate aminotransferase
(glutamate-oxaloacetate transaminase)

NH2 Aspartate O Oxaloacetate

HOOC-CH-CH2COOH HOOC-C-CH2COOH
+ glutamate
+ α-ketoglutarate

Alanine aminotransferase
(glutamate-pyruvate transaminase)

NH2 Alanine O Pyruvate

HOOC-CH-CH3 HOOC-C-CH3
+ glutamate
+ α-ketoglutarate

Figure 3. The reactions catalyzed by aspartate aminotransferase and

alanine aminotransferase.
 Glutamate
dehydrogenase

NADH NAD+

Glutamate
α-Ketoglutarate +
NH4+ NH3 + ATP
Glutamine
synthetase

ADP + Pi
Glutamine

Figure 3. In non-hepatic tissues the linked reactions of glutamate

dehydrogenase and glutamine synthetase remove two ammonia
molecules from the tissues as a way of ridding the tissues of nitrogen
waste. The glutamine deposits the ammonia in the kidney for excretion.
 Glutamine
Glutaminase
NAD+ NADH
NH4+
α-Ketoglutarate
Glutamate
+ NH4+
Glutamate dehydrogenase

Figure 5. Kidney production of ammonia for excretion following

successive removal of amino groups from glutamine via glutaminase
and glutamate dehydrogenase
α-Amino acid α-Ketoglutarate NADH + NH
NH44++
Urea
cycle
Aminotransferase Glu dehydrogenase
UREA

α-Keto acid
Glutamate NAD+ + H2O

Figure 6. In liver, nitrogen waste from amino acids ends up in urea.

Amino acids are derived either from the breakdown of protein in various
tissues or from what is synthesized in those tissues
 Fumarate O
(returns to UREA
2ATP + HCO3- + NH4+
TCA H2N-C- NH2
cycle) Arginine Carbamoyl
 phosphate
  synthetase
Argininosuccinate Ornithine 2ADP + Pi
Ornithine
AMP+PPi Carbamoyl phosphate

Citrulline
 Ornithine
transcarbamoylase

Citrulline Pi
Aspartate
-
ATP
-
OOC-CH-NH3+
CYTOPLASM MITOCHONDRIA

CH2COO-

 argininosuccinate synthetase  argininosuccinase arginase

Figure 7. Carbamoyl phosphate synthetase reaction and the urea cycle.

Overall: 3ATP+HCO3-+NH4++asp  2ADP+AMP+2Pi+PPi+fumarate+urea
 UREA CYCLE FACTS

 Found primarily in liver and lesser extent in kidney

 Nitrogen added to the urea cycle via carbamoyl phosphate and

aspartate

 Carbamoyl phosphate synthetase is allosterically activated by

N-acetylglutamate
(acetyl CoA + glutamate → N-acetylglutamate)

 Arginine stimulates the formation of N-acetylglutamate

 HYPERAMMONEMIAS
Acquired = Liver disease leads to portal-systemic shunting

Inherited = Urea cycle enzyme defects of CPS I or ornithine

transcarbamoylase lead to severe hyperammonemia

Fatty liver can lead to cirrhosis

 primary defect in Phenylalanine
phenylketonuria hydroxylase
O2 H2O
Phenylpyruvate Phenylalanine
X Tyrosine

Tetrahydrobiopterin Dihydrobiopterin
Phenyllactate

Phenylacetate

NADP+ NADPH

Figure 8. Unusual compounds produced from phenylalanine in

phenylketonuria. The phenylalanine hydroxylase reaction (or
regeneration of the tetrahydrobiopterin cofactor) are defective in
phenylketonuria.