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 Argininea Histidine Isoleucine Leucine Lysine
a

Nonessential Alanine Aspartate Asparagine Cysteine Glutamate Glutamine Glycine Proline Serine Tyrosine

Methionineb Phenylalaninec Threonine Tryptophan Valine

Arg is synthesized in the urea cycle, but the rate is too slow to meet the needs of growth in children Met is required to produce cysteine if the latter is not supplied adequately by the diet. supplied

b

c

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

BODY PROTEIN Proteolysis Carbon compounds + nitrogen Dietary amino acids De novo synthesis Amino Acid Pool Protein synthesis Catabolism

Urea + CO2

Biosynthesis of 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 HOOC-CH-R α-Amino acid

O HOOC-C-CH2CH2COOH α-Ketoglutarate Cofactor = pyridoxal phosphate

α-Keto acid O HOOC-C-R

Glutamate NH2 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-C-CH2COOH + glutamate

HOOC-CH-CH2COOH + α-ketoglutarate

Alanine aminotransferase (glutamate-pyruvate transaminase) NH2 Alanine O Pyruvate HOOC-C-CH3 + glutamate

HOOC-CH-CH3 + α-ketoglutarate

Figure 3. The reactions catalyzed by aspartate aminotransferase and alanine aminotransferase.

Glutamate dehydrogenase NADH α-Ketoglutarate + NH4+ NAD+ Glutamate 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+ NH4+ Glutamate Glutamate dehydrogenase α-Ketoglutarate + NH4+ NADH

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 + NH44++ NH

Urea cycle UREA

Aminotransferase

Glu dehydrogenase

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 (returns to TCA cycle)

O

UREA

H2N-C- NH2 Arginine  

2ATP + HCO3- + NH4+


Ornithine Citrulline Ornithine

Carbamoyl phosphate synthetase 2ADP + Pi

Argininosuccinate AMP+PPi


-

Aspartate OOC-CH-NH3+
-

Carbamoyl phosphate  Ornithine transcarbamoylase Citrulline Pi MITOCHONDRIA

ATP

CYTOPLASM

 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 aspartate and

 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 phenylketonuria

Phenylalanine hydroxylase O2 H2O

Phenylpyruvate

Phenylalanine

X

Tyrosine Dihydrobiopterin

Phenyllactate Phenylacetate

Tetrahydrobiopterin

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.

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