[A kinetic study of the reactive capabilities of xanthine oxidase sulfhydryl groups with regard to n-chlormercuribenzoate].
Kinetic characteristics for reactivity of SH-groups of milk xanthine oxidase were obtained under different conditions. Two types of SH-groups with rate constant values, differing by a factor of about 50, were found in a phosphate buffer at pH 7.0. The slow stage of reaction is followed by protein precipitation. The number of fast- (12) and slowly-reacting (60) groups were calculated from the kinetic data. The blocking of the fast-reacting groups occurs without loss of the enzyme activity. The values of activation energy for the fast- and slowly-reacting groups are 15 and 48 kcal/mol respectively. The formation of the enzyme-substrate complex stabilizes the enzyme molecule; the number of fast-reacting SH-groups and the rate constant values for both types of groups remain unchanged, whereas the number of slowly-reacting SH-groups markedly decreases (37). The values of activation energy for both types of SH-groups show no changes in the presence of substrate. Conformations of the enzyme in different denaturating solvents were characterized by a number of SH-groups, reacting with p-chloromercurybenzoate. 54 groups are exposed in solutions of groups exposed in 7.0-8.5 M urea solutions is 35-38. In all solvents studied the protein molecule is probably not completely unfolded, since the number of exposed SH-groups is less than the full number of SH-groups determined by the amino acid analysis. Only 42 SH-groups reacted with 5,5'-dithiobis-(2-nitrobenzoic acid) under the same conditions.
D000818:Animals; D002384:Catalysis; D002417:Cattle; D002729:Chloromercuribenzoates; D007700:Kinetics; D008892:Milk / Q000201:enzymology*; D008968:Molecular Conformation; D011489:Protein Denaturation; D013438:Sulfhydryl Compounds; D014969:Xanthine Oxidase
D002729:Chloromercuribenzoates; D013438:Sulfhydryl Compounds; D014969:Xanthine Oxidase
Kineticheskoe issledovanie reaktsionnoĭ sposobnosti sul'fgidril'nykh grupp ksantinoksidazy po otnosheniiu k n-khlormercuribenzoatu