Thiol oxidase activity of copper, zinc superoxide dismutase.

The ability of copper,zinc superoxide dismutase (Cu,Zn-SOD) to catalyze autoxidation of cysteine and other thiols was investigated by measuring thiol loss and oxygen consumption. The reaction occurred equally well with the bovine and human enzymes and produced hydrogen peroxide and the corresponding disulfide. It did not occur with manganese SOD and is not, therefore, due to the dismutase activity of the enzyme. Cysteine and cysteamine were highly reactive: the K(m) for cysteine was 1.4 mm and V(max) (with 40 microg/ml SOD) 35 microm/min; the equivalent values for cysteamine (with 20 microg/ml SOD) were 1.4 mm and 36 microm/min. With 1 mm thiol and 40 microg/ml SOD, rates of oxidation of other thiols (microm/min) were as follows: GSH, 1.0; dithiothreitol, 2.1; dihydrolipoic acid, 1.7; homocysteine, 1.6; cys-gly, 1.4; penicillamine, 0.6; and N-acetylcysteine, 0.1. SOD-mediated oxidation of cysteine, in the absence of chelating agents, proceeded only after a variable lag phase. The lag was decreased but not eliminated with Chelex-treated reagents and is attributed to interference by submicromolar concentrations of iron and possibly other transition metal ions. SOD-catalyzed oxidation of the other thiols was variably affected by adventitious metal ions and chelating agents. Reactions were all performed in the presence of desferrioxamine to obviate these effects. SOD-catalyzed oxidation of GSH and homocysteine was enhanced by cysteine through a thiol-disulfide exchange mechanism. This study characterizes a novel pro-oxidant thiol oxidase activity of Cu,Zn-SOD. It is a potential source of reactive oxidants and may contribute to the cytotoxicity of reactive thiols such as cysteine and cysteamine.