title: Mitochondrial GTP-AMP phosphotransferase. 2. Kinetic and equilibrium dialysis studies.
abstract: Kinetic and equilibrium dialysis substrate binding studies have been done to investigate the properties of mitochondrial GTP-AMP phosphotransferase. The results show that the enzyme has a specific requirement for divalent metal ions, namely Mg2+, Mn2+ or Ca2+ (Ca2+ is active only in the forward direction, the direction of formation of ADP). The reaction rate depends upon the ratio [Mg2+]:[substrate] rather than on the metal ion concentration alone. The enzymatic activity is influenced by NaCl (or KCl) and optimum pH occurs at 11.5 and 9.5 for guanosine and inosine nucleotides respectively. Examination of binding of substrates to the enzyme showed that there is one binding site (GTP site) for MgGTP, GTP, MgGDP or GDP per molecule of enzyme, with dissociation constants of 4.5, 4.4, 3.0, 2.2 micron respectively and one binding site (AMP site) for AMP, ADP or ATP per molecule of enzyme with dissociation constants of 20.9, 33.4 and 33.4 microns respectively. Since, within the limitations of equilibrium dialysis used in the present studies, AMP binding to one site of the enzyme could be detected only when GDP or GTP is present, the mechanism of the forward reaction may be assumed to be nearly ordered. For the reverse reaction there is no requirement of order of binding of the two nucleotides and so the mechanism of reaction may be assumed to be random.
mesh_terms: D000249:Adenosine Monophosphate; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D003956:Dialysis; D006160:Guanosine Triphosphate; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D009703:Nucleoside-Phosphate Kinase; D009994:Osmolar Concentration; D010770:Phosphotransferases / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D002413:Cations, Divalent; D000249:Adenosine Monophosphate; D006160:Guanosine Triphosphate; D010770:Phosphotransferases; C021894:nucleoside triphosphate-adenylate kinase; D009703:Nucleoside-Phosphate Kinase