UREA CYCLE Urea is the end product of nitrogen catabolism in humans Introduction: A human subjects who consumes 300

g of carbohydrate and 100 g of fat and 100 g of protein ,daily excretes about 16.5 g of nitrogen each day ,95% in the urine and 5% in the feces .For subjects consuming occidental diets ,urea synthesized in the liver ,released in to the blood and cleared by the kidneys ,constitutes 80to 90% of the nitrogen excreted . BIOMEDICAL IMPORTANCE: Deficiency of urea cycle enzymes and liver diseases the urea cycle is impaired and cause hyper ammonemia.

Pathway: Synthesis of 1 molecule of urea requires 3 molecules of ATP and 1 molecule of each ammonium ion and the alpha amino nitrogen of aspartate. Five enzymes catalyze the numbered reactions. Of the six participating amino acids, N acetyl glutamate functions solely as an enzyme activator. The other serves as carriers of atoms that ultimately become urea. In mammals, the major metabolic role of ornithine, citrulline and argino succinate is urea synthesis. Urea synthesis is a cyclic process, ornithine consumed in a reaction 2 is regenerated in reaction 5and there is no net loss or gain of ornithine, citrulline, arginino succinate or arginine. Ammonium ion, carbon dioxide, ATP and Aspartate are however consumed.

Occurrence: Some reactions of urea synthesis occur in the mitochondrion while others occur in the cytosol. REACTIONS INVOLVED IN UREA CYCLE: 1. Carbamoyl phosphate synthase I : The biosynthesis of urea begins with the condensation of carbon dioxide; ammonia and ATP to form carbamoyl phosphate, a reaction catalysed by carbamoyl phosphate synthase I. Human tissue contain two forms of carbamoyl phosphate synthase. CPS-I, the enzyme functional in urea synthesis, is hepatic mitochondrial enzyme, carbamoyl phosphate synthase II is cytosolic enzyme that uses glutamine rather than ammonia as the nitrogen donor, functions in pyrimidine biosynthesis. Formation of carbamoyl phosphate requires 2 moles of ATP. One ATP serves as a source of phosphate. Conversion of the second ATP to AMP and pyrophosphate together with the coupled hydrolysis of pyrophosphate to ortho phosphate provides the driving force for the synthesis of amide bond and the mixed anhydride bond of carbamoyl phosphate.

CPS I is the rate limiting or pace maker enzyme of the urea cycle, this regulatory enzyme is active only in the presence of the allosteric activator N acetyl glutamate, whose binding induces a conformational change that enhances the affinity of the synthetase for ATP. 2. Ornithine Trans carbamoylase ( O- T ) It catalyses the transfer of carbamoyl moiety of carbamoyl phosphate to ornithine, forming citrulline plus ortho phosphate. While this reaction is takes place in the mitochondrial matrix, the compartment in which the substrate ornithine is formed and product citrulline is further metabolized in the cytosol. Entry of ornithine into mitochondria and exodus of citrulline from mitochondria therefore involve mitochondrial inner membrane transport system (anti citrulline ornithine transporter) 3. Arginino Succcinate synthase: The reaction links aspartate and citrulline via the amino group of aspartate and provides the second nitrogen of urea. The reaction requires ATP and involves the intermediate formation of citrullyl AMP. Subsequent displacement of AMP by aspartate then forms citrulline. 4. Arginino Succinase: Cleavage of arginino succinate, a reversible Trans elimination reaction catalyzed by arginino succinase, retains nitrogen in the product arginine and releases aspartate skeleton as fumarate. Addition of water to fumarate forms L malate and subsequent NAD dependent oxidation of malate forms oxalo acetate. 5. Arginase: The final reaction of urea cycle, the hydrolytic cleavage of guanidino group of arginine is catalyzed by liver arginase releases urea. The other product ornithine reenters liver mitochondria for the additional rounds of the urea cycle. Ornithine and lysine are potent competitive inhibitors of arginase REGULATION: The activity of carbamoyl phosphate synthase I is determined by the steady state concentration of its allosteric activator N-acetyl glutamate. This steady state level is determined by the rates of its synthesis from acetyl coA and glutamate and hydrolysis to acetate and glutamate, reactions catalysed by N acetyl glutamate synthase and N-acetyl glutamate hydrolase respectively. Metabolic disorders associated with each reaction of the urea cycle: Urea synthesis converts toxic ammonia to non toxic urea, all defects in urea synthesis result in ammonia intoxication. This intoxication is more severe when the metabolic block occurs at reactions 1 or 2, since some covalent linking of ammonia to carbon as already occurred if citrulline can be synthesized. Clinical symptoms include vomiting, avoidance of high protein food, intermittent ataxia, irritability, lethargy and mental retardation. 1. Hyper ammonemia I

Deficiency of carbamoyl phosphate synthease I 2. Hyper ammonemia II Deficiency of ornithine trans carbamoylase have symptoms that includes elevation of glutamine in blood, CSF and urine, reflects enhanced glutamine synthesis, consequent to elevated tissue levels of ammonia 3. citrullinemia: Nearly 1 to 2 g of citrulline is excreted in the urine daily, and both plasma and CSF citrulline levels are elevated due to the deficiency of arginino succinate activity. Note: Feeding benzoate diverts ammonia nitrogen to hippurate via glycine 4. Arginino succinic aciduria: Characterized by elevated levels of arginino succinate in the blood and CSF and urine. It reflects the absence of arginino succinase Note: feeding arginine and benzoate promotes nitrogen excretion. 5. Hyper argininemia: Characterized by elevated blood and CSF arginine levels, low RBC levels of arginase and urinary amino acid pattern resembling that of lysine cystinuria.