Chromium (V) and Hydroxyl Radical Formation during the Glutathione Reductase-Catalyzed Reduction of Chromium (VI)

"Electron spin resonance measurements provide evidence for the formation of long-lived Cr(V) intermediates in the reduction of Cr(Vl) by glutathione redtuctase in the presence of NADPH and for the hydroxyl radical formation during the glutathione reductase catalyzed reduction of Cr(Vl). Hydrogen peroxide suppresses Cr(V) and enhances the formation of hydroxyl radicals. Thus Cr(V) intermediates catalyze generation of hydroxyl radicals from hydrogen peroxide through a Fenton-like reaction. Thus the mechanism of Cr(VI) toxicity might involve the interaction between macromolecules and the hydroxyl radicals.C(Vl) compounds have been found to exert serious toxic and carcinogenic effects on humans and animals (1-3). In contrast, most Cr(III) compounds are relatively nontoxic, noncarcinogenic, and nonmutagenic (1,4). Cr(VI) and Cr(III) oxidation states are different in their metabolic pathways: Cr(VI) , ions are rapidly transported across cellular membrane (1,4,5), while Cr[III) moieties do not easily penetrate cells and are not oxidized by cellular constituents (!). Since it is known that the reduction of Cr(VI) ion is required for its reaction with DNA (1), the molecular mechanisms for the intracellular Cr(Vl) reduction has been the focus of current studies but the details are still not understood.Earlier studies on the mechanism of Cr(Vl) reduction include those on the reduction of Cr(VIJ by glutathione (GSH) .0,6-8), ascorbic acid (9), glucose (10), lactose (IO), galacturonic acid (11), microsome (12), and mitochondrial electron transport chain complexes (13). These studies have shown that the Cr(VI) reduction involves the formation of Cr(V)-containing species which are often considered to be the toxic form of chromium. Thus far, however, no information is· available on the subsequent fate of those biologically formed Cr(V) species, in particular employing well defined biochemical systems. In this context, we report here our investigations of the reduction of Cr(VI) by glutathione reductase (GSSG-R). This enzyme was selected because it is ubiquitously present in cellular systems and it has been postulated to be involved in the Cr(VI) reduction (14-17) but the underlying mechanism has not been clarified. We find that the reduction proces·s involves not only Cr(V) but also ·OH radicals. We further investigate the reactivity of Cr(V) and find that it catalyzes generation of OH radical from H2O 2 through a Fenton-like reaction. The role of ·OH in Cr(VI) carcinogenicity is suggested from this study."