Inactivation of glucose oxidase by diperoxovanadate-derived oxidants.
Inactivation of glucose oxidase occurred in the presence of bromide, vanadate, H(2)O(2), and phosphate (the bromide system), and this was prevented by NADH or phenol red, a bromine acceptor. Glucose oxidase present during the reaction between diperoxovanadate and a reduced form of vanadate, vanadyl (the vanadyl system), but not added after mixing the reactants, was inactivated, and this was accompanied by a loss of binding of the dye, Coomassie blue, to the protein. The transient intermediate of the type OVOOV(O(2)), known to form in these reactions and used in the oxidation of bromide ion and NADH, appears to be responsible for inactivating glucose oxidase. In both systems, the inactivation of the enzyme was prevented by histidine and DTT, known to quench singlet-oxygen. By direct measurement of 1270-nm emission of singlet-oxygen, its generation was demonstrated in the bromide system, and in the reaction of hypohalous acids with diperoxovanadate, but not in the vanadyl system. By themselves both hypohalous acids, HOCl and HOBr inactivated glucose oxidase, and their prior reaction with H(2)O(2) during which singlet-oxygen was released, protected the enzyme. The results provide support for possible oxidative inactivation of glucose oxidase by diperoxovanadate-derived oxidants.
Rao
A V
AV
Department of Biochemistry, Indian Institute of Science, Bangalore, 560012, India.
Sima
P D
PD
Kanofsky
J R
JR
Ramasarma
T
T
D001965:Bromides / Q000494:pharmacology; D005949:Glucose Oxidase / Q000037:antagonists & inhibitors*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007700:Kinetics; D008956:Models, Chemical; D016877:Oxidants / Q000494:pharmacology*; D010545:Peroxides / Q000737:chemistry* / Q000494:pharmacology*; D013481:Superoxides / Q000378:metabolism; D014638:Vanadates / Q000737:chemistry* / Q000494:pharmacology*
D001965:Bromides; D016877:Oxidants; D010545:Peroxides; C091767:diperoxovanadate; D013481:Superoxides; D014638:Vanadates; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase