title: Kinetics of inactivation of creatine kinase during modification of its thiol groups.
abstract: Kinetics of inactivation and modification of the reactive thiol groups of creatine kinase by 5,5'-dithiobis(2-nitrobenzoic acid) or iodoacetamide have been compared, the former by following the substrate reaction in presence of the inactivator [Wang, Z.-X., & Tsou, C.-L. (1987) J. Theor. Biol. 127, 253]. The microscopic constants for the reaction of the inactivators with the free enzyme and with the enzyme-substrate complexes were determined. From the results obtained it appears that with respect to ATP both inactivators are noncompetitive whereas for creatine iodoacetamide is competitive but DTNB is not. The formation of the ternary complex protects against the inactivation by both DTNB and iodoacetamide. The inactivation kinetics is monophasic with both inactivators, but under similar conditions, the modification reactions in the presence of the transition-state analogue of creatine-ADP-Mg2+-nitrate show biphasic kinetics as also reported by Price and Hunter [Price, N.C., & Hunter, M.G. (1976) Biochim. Biophys. Acta 445, 364]. If the reactive ternary complex and the enzyme complexed with the transition-state analogue react in the same way with these reagents, the modification of one fast-reacting thiol group for each enzyme molecule leads to complete inactivation, indicating that the enzyme has to be in the dimeric state to be active.
authors.affiliation: Laboratory of Molecular Enzymology, Institute of Biophysics, Academia Sinica, Beijing, China.
authors.forename: Z X
authors.initials: ZX
authors.lastname: Wang
authors.forename: B
authors.initials: B
authors.lastname: Preiss
authors.forename: C L
authors.initials: CL
authors.lastname: Tsou
mesh_terms: D000818:Animals; D003402:Creatine Kinase / Q000037:antagonists & inhibitors*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology; D007460:Iodoacetamide / Q000494:pharmacology; D007700:Kinetics; D008433:Mathematics; D009132:Muscles / Q000201:enzymology; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D013438:Sulfhydryl Compounds; D004228:Dithionitrobenzoic Acid; D003402:Creatine Kinase; D007460:Iodoacetamide