NADPH-dependent Flavoenzymes Catalyze One Electron Reduction of Metal Ions and Molecular Oxygen and Generate Hydroxyl Radical

"This study reports a new property of the important NAD(P)H-dependent flavoenzymes, glutathione reductase, lipoyl dehydrogenase and ferredoxin- NADP oxidoreductase, that can catalyze a one electron reduction of metal ions such as chromium(VI) and vanadium(V). During the enzymatic reduction process, molecular oxygen is reduced to H2O 2 , which reacts with the reduced metal complexes to generate hydroxyl radicals. Since the hydroxyl radicals have been suggested to play an important role in Cr(VI) toxicity, this study provides a basis for a recent observation that Cr(VI) mutagenesis is strongly oxygen dependent. These results also point to an enzymatic pathway for the metabolism of some metal ions and concomitant generation of hydroxyl radicals. l. INTRODUCTIONThe purpose of this communication is to report that some important NAD(P)H-dependent flavoenzymes (glutathione reductase, lipoyl dehydrogenase and ferredoxin-NADP+ oxidoreductase) can catalyze one electron reduction of metal ions such as chromium(VI) and vanadium(V) and simultaneously reduce molecular oxygen to generate Hi02 and, eventually, the ""OH radical. This finding is significant because (i) the known substrates for the flavoenzymes are organic species and not metal ions, and (ii) the report on the enzymatic formation of the ""OH radical provides a new pathway for understanding the mechanism of genotoxicity of some non-ferrous metal ions. For example, it has been suggested that OH radical is likely to be the major species responsible for Cr(VI) genotoxicity [1-5). The 'OH radical is generated through the reaction of Cr(V) complexes with cellular Hi02 [4,5]. Cr(V) itself is known to be generated from Cr(VI) in various biological systems, in particular, microsomes [ 6), mitochondria [7] and mitochondrial electron transfer chain complexes [8]. However, no specific enzyme responsible for the Cr(V) generation has been identified except possibly glutathione reductase [4]. In this study we have used ESR and spin trapping techniques to show that the flavoenzymes can generate Cr(V) and provide a catalytic redox cycle for 'OH formation in such reactions. Herein we present our results on Cr(VI) and summarize those on other systems. Correspondence address: N.S."