Oxidation of catechol by horseradish peroxidase and human leukocyte peroxidase: Reactions of o-benzoquinone and o-benzosemiquinone

Abstract

The metabolism of secondary phenolic metabolites of benzene, such as catechol, by peroxidases represents one possible mechanism underlying benzene-induced myelotoxicity. The oxidation of catechol by horseradish peroxidase and peroxidases present in human leukocytes was therefore examined. Peroxidatic oxidation resulted in o-benzoquinone production, which was characterized as its bromothiophenol adduct. o-Benzoquinone-glutathione conjugates were formed during peroxidatic oxidation of catechol in the presence of glutathione. Both mono- and diglutathione conjugates were detected. As much as 80% of catechol removed during peroxidatic oxidation could be recovered as glutathione conjugates of o-benzoquinone. Glutathione had no inhibitory effect on the removal of catechol during peroxidatic oxidation. In the presence of divalent cations (Mg²⁺, Zn²⁺), however, which slow the rate of o-semiquinone disproportionation, glutathione was found to inhibit catechol removal. This suggests that in the absence of stabilizing metal, reduction of the o-benzosemiquinone radical by glutathione cannot compete with other rapid reactions of the radical such as disproportionation. No interaction of the o-benzosemiquinone radical with oxygen could be detected even in the presence of stabilizing metals or superoxide dismutase which inhibits the reverse reaction of the $\text{S}\text{̇}\text{Q + O}{}_2\text{α Q +}\text{O}\text{·}{}_2{}^{\mathrm{•}}$ equilibrium. Thus, under physiological conditions, glutathione and oxygen would not be expected to reduce or oxidize respectively the o-benzosemiquinone radical. These data show that the generation of thiol conjugates of o-benzoquinone can be used as probes of peroxidatic oxidation of catechol.