title: [Effect of pH on conformation and kinetic properties of phosphorylase from bovine skeletal muscles].
abstract: Interaction of phosphorylase with 8-anilino-1-naphthalene-sulfonate (ANS) results in the formation of an ANS-protein complex. The microenvironment of the protein-bound dye changes depending on pH. Using fluorimetric titration, the dissociation constants for the complex (Kd = 23 and 57 microM for pH 6.2 and 6.8, respectively) were determined. The mode of the enzyme inhibition by ANS also changes depending on pH. At pH 6.8, ANS competitively inhibits the enzyme with respect to AMP, but does not compete with the nucleotide at pH 6.2; the corresponding Ki values are equal to 160 and 26 microM. The protective effect of ligands from the inhibiting effect of ANS was studied. It was shown that at pH 6.2, the enzyme is protected from the inhibition only by the substrate, glucose-1-phosphate, whereas at pH 6.8--by the allosteric inhibitor, glucose-6-phosphate. These findings suggest that at pH 6.2 the conformation of the enzyme molecule is induced by the substrate, while at pH 6.8--by the allosteric inhibitor. ANS binding in the vicinity of the active or allosteric centers is due to the pH-dependent conformational transition. The data obtained suggest that the pH changes within the range of 6.2-6.8 are essential for the regulation of enzyme activity.
authors.forename: L K
authors.initials: LK
authors.lastname: Skolysheva
authors.forename: S A
authors.initials: SA
authors.lastname: Shur
authors.forename: P L
authors.initials: PL
authors.lastname: Vul'fson
mesh_terms: D000817:Anilino Naphthalenesulfonates; D000818:Animals; D002417:Cattle; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D006005:Phosphorylases / Q000032:analysis* / Q000037:antagonists & inhibitors; D011487:Protein Conformation; D011817:Rabbits; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; C027132:1-anilino-8-naphthalenesulfonate; D006005:Phosphorylases
vernacular_title: Vliianie pH na konformatsionnye i kineticheskie svoĭstva fosforilazy iz skeletnykh myshts byka.