Hepatic carbamoyl phosphate metabolism. Role of cytosolic and mitochondrial carbamoyl phosphate in de novo pyrimidine synthesis.
The interrelationship between the two carbamoyl phosphate pools in intact hepatocytes and intact liver was studied with respect to de novo pyrimidine synthesis by use of selective inhibitors of the mitochondrial and the cytosolic carbamoyl-phosphate synthetase. Inhibition of mitochondrial carbamoyl phosphate synthesis by 4-pentenoate was without effect on galactosamine-stimulated pyrimidine synthesis. Conditions favouring mitochondrial carbamoyl phosphate accumulation, like excess ammonium ions or L-norvaline, led to an increase in pyrimidine synthesis bypassing the feedback inhibition of cytosolic carbamoyl-phosphate synthetase by UTP. A stimulation of pyrimidine synthesis was not observed when the carbamoyl phosphate accumulation was due to aspartate deficiency in the presence of aminooxyacetate. The full response of pyrimidine synthesis to excess ammonium ions was restored, even in the presence of aminooxyacetate, when aspartate was substituted. This is explained by an inhibition of aspartate carbamoyltransferase flux [in view of the Km (aspartate = 0.7 mmol/l) of this enzyme] resulting from a 90% decrease in aspartate tissue levels. Acivicin, the inhibitor of cytosolic carbamoyl-phosphate synthetase, completely abolished the galactosamine-induced stimulation of pyrimidine synthesis, but was without effect on the stimulation of pyrimidine synthesis by ammonium ions and L-norvaline. It is concluded that experimental changes in mitochondrial carbamoyl phosphate content exert effects on de novo pyrimidine synthesis; however, it is considered unlikely that relevant amounts of mitochondrial carbamoyl phosphate are used for pyrimidine synthesis under physiological conditions. In addition the data point to a potential regulatory role of aspartate in hepatic pyrimidine synthesis.
D000625:Aminooxyacetic Acid / Q000494:pharmacology; D000818:Animals; D002219:Carbamates / Q000378:metabolism*; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / Q000037:antagonists & inhibitors; D002221:Carbamyl Phosphate / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D007555:Isoxazoles / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism; D009954:Ornithine Carbamoyltransferase / Q000037:antagonists & inhibitors; D009963:Orotic Acid / Q000378:metabolism; D010477:Perfusion; D011743:Pyrimidines / Q000096:biosynthesis*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D000637:Transaminases / Q000037:antagonists & inhibitors; D014633:Valine / Q000494:pharmacology / Q000031:analogs & derivatives
D002219:Carbamates; D005229:Fatty Acids, Monounsaturated; D005231:Fatty Acids, Unsaturated; D007555:Isoxazoles; D011743:Pyrimidines; D000644:Quaternary Ammonium Compounds; D000625:Aminooxyacetic Acid; D002221:Carbamyl Phosphate; D009963:Orotic Acid; C005313:norvaline; C009785:4-pentenoic acid; D009954:Ornithine Carbamoyltransferase; D000637:Transaminases; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing); D014633:Valine; C006303:acivicin