Studies of the regulation and reaction mechanism of the carbamyl phosphate synthetase and aspartate transcarbamylase of bakers' yeast.
Kinetic studies of the carbamyl phosphate synthetase activity (CPSase) of bakers' yeast revealed an absolute requirement for K+ ions ; KM values for two of the substrates, glutamine and bicarbonate, were found to be 5 X 10(-4) M and 3 X 10(-3) M respectively. CPSase activity of the purified enzyme aggregate (M.W. 800,000) was extremely sensitive to UTP with a Ki of 2.4 X 10(-4) M. The purine nucleotide intermediate, XMP, was a strong activator of CPSase, acting at a site different from the regulatory site at which UTP binds ; XMP activation diminished at high concentrations of the substrate Mg-ATP. Studies of the reaction mechanism of CPSase revealed that it involved the sequential addition of the substrates bicarbonate and Mg-ATP, liberation of ADP, addition of glutamine, binding of ATP and then release of ADP and the product carbamyl phosphate. Studies of the reaction mechanism of the aspartate transcarbamylase (ATCase) of the aggregate yielded data which were not compatible with any of the usual models ; whichever reaction mechanism is ultivately found to fit the data, it will probably prove applicable both to the ATCase of the aggregate and to the disaggregated ATCase subunit (MW 138,000).
D000255:Adenosine Triphosphate / Q000378:metabolism; D001221:Aspartate Carbamoyltransferase / Q000378:metabolism*; D001639:Bicarbonates / Q000378:metabolism; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005973:Glutamine / Q000378:metabolism; D007700:Kinetics; D008274:Magnesium; D010770:Phosphotransferases / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D011685:Purine Nucleotides / Q000494:pharmacology; D012265:Ribonucleotides / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D014500:Uracil Nucleotides / Q000494:pharmacology; D014970:Xanthines / Q000494:pharmacology
D001639:Bicarbonates; D011685:Purine Nucleotides; D012265:Ribonucleotides; D014500:Uracil Nucleotides; D014970:Xanthines; D005973:Glutamine; D000255:Adenosine Triphosphate; D001221:Aspartate Carbamoyltransferase; D010770:Phosphotransferases; D002222:Carbamoyl-Phosphate Synthase (Ammonia); D008274:Magnesium; D011188:Potassium