title: A differential scanning calorimetric study of the binding of sulfate ion and of Cibacron blue F3GA to yeast phosphoglycerate kinase.
abstract: In continuation of earlier work [Hu, C. Q., & Sturtevant, J.M. (1987) Biochemistry 26, 178-182], differential scanning calorimetry has been employed in a study of the effects on the thermal denaturation of yeast phosphoglycerate kinase of two inhibitors of the enzyme, sulfate ion and the dye Cibracron blue F3GA. Sulfate ion, as is usual with ligands that dissociate during unfolding of the host protein, raises t1/2, the temperature of half-completion of the denaturation, has only a modest effect, stemming from the enthalpy of dissociation of the ligand, on the enthalpy of denaturation, and has little or no effect on the heat capacity change resulting from denaturation. In sharp contrast, Cibacron blue F3GA lowers t1/2 and drastically decreases both the enthalpy and heat capacity changes due to denaturation. The DSC results with sulfate ion are consistent with previous kinetic data [Scopes, R. K. (1978) Eur. J. Biochem. 91, 119-129; Khamis, M. H., & Larsson-Raznikiewicz, M. (1981) Biochim. Biophys. Acta 657, 190-194], which indicate two binding sites for sulfate ion at one of which the ligand acts as a competitive inhibitor. The results with Cibacron blue F3GA indicate that the dye induces a major destabilizing structural change in the enzyme in addition to rendering it enzymically inactive.
authors.affiliation: Department of Chemistry, Yale University, New Haven, Connecticut 06511.
authors.forename: C Q
authors.initials: CQ
authors.lastname: Hu
authors.forename: J M
authors.initials: JM
authors.lastname: Sturtevant
mesh_terms: D002152:Calorimetry, Differential Scanning; D004396:Coloring Agents; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013431:Sulfates / Q000378:metabolism*; D014227:Triazines / Q000378:metabolism*
chemical_list: D004396:Coloring Agents; D013431:Sulfates; D014227:Triazines; C012842:Cibacron Blue F 3GA; D010735:Phosphoglycerate Kinase