On the Mechanism of the Chromate Reduction by Glutathioe: ESR Evidence for the Glutatbionyl Radical and an Isolable Cr(V) Intermediate

"With a view of elucidating the role of glutathione (GSH) in the biochemical pathways of the chromate-exposure related carcinogenesis, we carried out electron spin resonance (ESR) spectroscopic investigations of the chromate-GSH redox reactions. The ESR measurements, employing spin-traps, provide evidence for the involvement of the glutathione (GS•) radical, as well as an isolable Cr(V)-glutathione intermediate. These results indicater a new mechanism for the reduction of chromate by GSH in cellular environment and help understand the (unexpected increase in Cr(VI)-induced DNA strand breaks at elevated GSH levels.We report here indirect electron spin resonance (EGR evidence for the involvement of the glutathionyl (GS) radical and direct evidence for a Cr(V) intermediate in the reactions of glutathione ¬(GSH) with chromate for the first time.. This work was undertaken to clarify the biochemical pathway of the reduction of chromate by GSH, since this reduction process is thought to be a critical step in the pathogenesis of the chromate-exposure related carcinogenesis. This follows from the literature reports which show that (a) Cr(VI)compounds have been proven to be carcinogens in laboratory animals as well as in human studies2-4, (b) Cr(VI) and not Cr(III) compounds are mutagenic 5-7, (c) Cr(VI) and not Cr(III) compounds can penetrate cell membranes 8.9 (d) the final state of Cr(VI) in cellular reductions is Cr(III)', (e) by itself chromate (or Cr(VI)) cannot be directly mutagenic since Cr(VI) is known not to interact with isolated DNA under physiological conditions 10, suggesting the significance of Cr(VI).reduction processes. One of the major reductants in cellular environments is thought to be the GSH, both outside and inside the ce11s 11-13. Other evidence for the role of GSH in the chromate toxicity is seen from recent studies showing that exposure of hamster cells to non-toxic levels of added selenite increases the levels' of GSH as well as the Cr(VI)-induced DNA strand beaks 14, and thatsuch DNA strand breaks in hepatocytes also change in direct proportion to their GSH content 15,16. These observations wereinterpreted as implying that the reduction of chromate by GSH, to some reactive intermediate is an important step in the chromatecarcinogenicity 15,16. The following reaction steps were,suggested 15, 16"