Evidence against transition metal-independent hydroxyl radical generation by xanthine oxidase.
It is shown by the use of EPR spectroscopy that formation of the hydroxyl radical adduct with the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) in the xanthine-xanthine oxidase system is hydrogen peroxide-independent. Production of the DMPO-hydroxyl radical adduct is inhibited by superoxide dismutase but is unaffected by purified grades of catalase. Hydroxyl radicals are a secondary product of the decomposition of the DMPO-superoxide radical adduct and are also formed as a result of trace metals such as iron present in the buffer. These results are in contrast with a recent report (Kuppusamy, P., and Zweier, J. W. (1989) J. Biol. Chem. 264, 9880-9884) in which the assertion is made that the hydroxyl radical adduct arises from the trapping of hydroxyl radicals generated via the direct reduction of hydrogen peroxide by xanthine oxidase. It is demonstrated here that treatment of phosphate buffer with the chelator deferoxamine mesylate is not in itself sufficient to suppress the effect of contaminating adventitious metal ions in xanthine-xanthine oxidase incubations.
Lloyd
R V
RV
Laboratory of Molecular Biophysics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
Mason
R P
RP
D002374:Catalase / Q000378:metabolism / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D005609:Free Radicals; D006878:Hydroxides / Q000378:metabolism*; D017665:Hydroxyl Radical; D007700:Kinetics; D008670:Metals / Q000494:pharmacology*; D013113:Spin Labels; D013482:Superoxide Dismutase / Q000378:metabolism / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism*
D005609:Free Radicals; D006878:Hydroxides; D008670:Metals; D013113:Spin Labels; D013481:Superoxides; D017665:Hydroxyl Radical; D002374:Catalase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D003676:Deferoxamine