title: Evidence for allosteric regulation of succinyl-CoA synthetase.
abstract: We have previously reported that distinctly different concentrations of GDP stimulate the phosphorylation and dephosphorylation of p36, the alpha-subunit of succinyl-CoA synthetase (SCS) in Dictyostelium discoideum. In this present study, we have investigated the mechanism underlying these dual effects of GDP. Dephosphorylation of p36 is induced by relatively high levels of GDP and is coincident with the formation of GTP. This indicates that, at high concentrations, GDP serves as a substrate of SCS. However, 100-fold lower concentrations of GDP, which do not bind to the catalytic site to induce SCS dephosphorylation, stimulate p36 phosphorylation. This stimulation is not diminished by dilution of the sample, and is retained during purification of the protein. Gel-filtration analyses indicate that SCS in our system behaves as a non-interacting alpha beta dimer, the hydrodynamic behaviour of which is not altered by the presence of added GDP. The data indicate that altered protein-protein interactions do not account for the stimulation of p36 phosphorylation by low GDP concentrations. We propose that GDP functions as an allosteric regulator of SCS, and experiments using guanosine 5'-[beta-thio]diphosphate (GDP[S]) are shown to distinguish further the allosteric and catalytic binding sites.
authors.affiliation: E.A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, MO 63104.
authors.forename: H D
authors.initials: HD
authors.lastname: Um
authors.forename: C
authors.initials: C
authors.lastname: Klein
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D001665:Binding Sites; D002850:Chromatography, Gel; D004023:Dictyostelium / Q000201:enzymology*; D006153:Guanosine Diphosphate / Q000378:metabolism / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000378:metabolism; D046911:Macromolecular Substances; D010766:Phosphorylation; D013389:Succinate-CoA Ligases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D046911:Macromolecular Substances; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; D013389:Succinate-CoA Ligases