Reverse and forward reactions of carbamyl phosphokinase from Streptococcus faecalis R. Participation of nucleotides and reaction mechanisms.
The participation of Mg complex of nucleoside diphosphates and nucleoside triphosphates in the reverse and forward reactions catalyzed by purified carbamyl phosphokinase (ATP : carbamate phosphotransferase, EC 2.7.2.2) of Streptococcus faecalis R, ATCC-8043 were studied. The results of initial velocity studies of approx. 1 mM free Mg2+ concentration have indicated that in the reverse reaction MgdADP was as effective a substrate as MgADP. The phosphoryl group transfer from carbamyl phosphate to MgGDP, MgCDP and MgUDP was also observed at relatively higher concentrations of the enzyme and respective magnesium nucleoside diphosphate. In the forward direction MgdATP was found to be as efficient a phosphate donor as MgATP. On the other hand, Mg complexes of GTP, CTP and UTP were ineffective even at higher concentrations of the enzyme and respective magnesium nucleoside triphosphate. Product inhibition studies carried out at non-inhibitory level of approx. 1 mM free Mg2+ concentration have revealed that the enzyme has two distinct sites, one for nucleoside diphosphate or nucleoside triphosphate and the other for carbamyl phosphate or carbamate, and its reaction with the substrates is of the random type. Further tests of numerical values for kinetic constants have indicated that they are partially consistent with the Haldane relationship which is characteristic of rapid equilibrium and random mechanism.
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D003565:Cytidine Diphosphate / Q000378:metabolism; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D013293:Enterococcus faecalis / Q000201:enzymology*; D006153:Guanosine Diphosphate / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism*; D017851:Phosphotransferases (Carboxyl Group Acceptor); D013379:Substrate Specificity; D014530:Uridine Diphosphate / Q000378:metabolism
D003838:Deoxyadenine Nucleotides; D009711:Nucleotides; D006153:Guanosine Diphosphate; C018891:deoxyadenosine diphosphate; D014530:Uridine Diphosphate; D000244:Adenosine Diphosphate; D003565:Cytidine Diphosphate; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D017851:Phosphotransferases (Carboxyl Group Acceptor); C019288:carbamate kinase; D008274:Magnesium; C026600:2'-deoxyadenosine triphosphate