One-Electron Reduction of Carcinogen Chromate by Microsomes, Mitochondria, and Escherichia coli: Identification of Cr(V) and •OH RadicalShi,

While Cr(VI) is a known carcinogen, the underlying biochemical mechanism is still not fully understood (1). In a pioneering study Jennette (2) utilized electron spin resonance (ESR) spectroscopy to demonstrate that a long-lived Cr(V) species is generated during the reduction of Cr(VI) by microsomes in the presence of NADPH. Later Wetterhahn and co-workers (3, 4) showed that incubation of chromate with isolated rat liver mitochondria causes the reduction of Cr(VI), as well as the formation of a Cr(V) species. Analogous Cr(V) species have also been detected in the reduction of Cr(VI) by other cellular systems such as Hanserulla polymorphared (5) and blood cells (6, 7). All these as well as other studies (8-12) have suggested that this Cr(V) species plays a significant role in the mechanism of Cr(VI) toxicity. Thus far, however, the chemical structure of this dominant Cr(V) species formed in cellular systems has not been identified, and this observation provided an impetus for the present investigation. Another reason for undertaking this study was that some recent reports (10-16) suggest that hydroxyl ('OH) radicals may also play an important role in the mechanism of Cr(VI) toxicity. In particular, it has been shown that OH radicals are formed in reactions between Cr(V) species and H2O2 (13-16), and that the radical yield depends on the chemical structure of the Cr(V) species (15). For example, while Cr(V)-diol complexes efficiently generate OH radicals, another Cr(V) species, the tetraperoxochromate (CrO3/8) ion, does not (15). Thus in the present study, we focused our attention on the following: (a) structural identification of the Cr(V) species generated in the reduction of Cr(VI) by subcellular as well as cellular systems, using microsomes, mitochondria, and Escherichia coli; (b) possible detection of OH radical in the same systems since there is no earlier report of chromium-mediated generation of 'OH radicals in a cellular system; (c) possible involvement of molecular oxygen in chromium-related OH radical generation since a recent report (17) indicated that Cr(VI) mutagenesis is strongly oxygen dependent; and (d) elucidation of the mechanism of OH radical formation in these systems.