Pyruvate carboxylase catalysis of phosphate transfer between carbamoyl phosphate and ADP.
In a reaction that is analogous to the phosphorylation of ADP from carboxyphosphate, pyruvate carboxylase catalyses the formation of ATP from carbamoyl phosphate and ADP at a rate that is about 0.3% of the pyruvate-carboxylation reaction and about 3% of the full reverse reaction. Acetyl-CoA stimulates the phosphorylation of ADP from carbamoyl phosphate but is not an essential requirement of the reaction. Mg2+ also stimulates the reaction, and in the range of Mg2+ concentrations considered the effect of V is much larger in the absence of acetyl-CoA than in its presence. Acetyl-CoA and Mg2+ may be acting in a co-operative way to stimulate the phosphorylation of ADP in a similar way to their effects on the pyruvate-carboxylation reaction. The phosphorylation of ADP by carbamoyl phosphate is also stimulated by the presence of biotin in the part of the active site where this reaction occurs, but again it is not absolutely required for the reaction to proceed. The pH profiles of the phosphorylation of ADP by carbamoyl phosphate indicate that there are at least two ionizable residues involved in the reaction, one of which probably has a role in the release of carbamate from the active site.
Attwood
P V
PV
Department of Biochemistry, University of Western Australia, Nedlands.
Graneri
B D
BD
D000105:Acetyl Coenzyme A / Q000494:pharmacology*; D000244:Adenosine Diphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001710:Biotin / Q000494:pharmacology*; D002221:Carbamyl Phosphate / Q000378:metabolism*; D002645:Chickens; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology*; D010071:Oxaloacetates / Q000378:metabolism; D010072:Oxamic Acid / Q000494:pharmacology; D010766:Phosphorylation; D011766:Pyruvate Carboxylase / Q000378:metabolism*; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D013379:Substrate Specificity
D010071:Oxaloacetates; D011773:Pyruvates; D002221:Carbamyl Phosphate; D000244:Adenosine Diphosphate; D001710:Biotin; D000105:Acetyl Coenzyme A; D019289:Pyruvic Acid; D011766:Pyruvate Carboxylase; D008274:Magnesium; D010072:Oxamic Acid