title: Inactivation of xanthine oxidase by hydrogen peroxide involves site-directed hydroxyl radical formation.
abstract: The mechanism of xanthine oxidase (XO) inactivation by hydrogen peroxide (H2O2) and its biologic significance are unclear. We found that addition of increasing concentrations of H2O2 progressively decreased xanthine oxidase activity in the presence but not the absence of xanthine in vitro. Inactivation of XO by H2O2 was also enhanced by anaerobic reduction of XO by xanthine. Inactivation of XO by H2O2 was accompanied by production of hydroxyl radical (.OH), measured as formation of formaldehyde from dimethylsulfoxide (DMSO). In contrast, addition of H2O2 to deflavo XO did not produce .OH. Inactivation of XO by H2O2 was decreased by simultaneous addition of the .OH scavenger, DMSO. However, inactivation of XO by H2O2 and formation of .OH were not decreased following addition of the metal chelator. DETAPAC, and/or the O2 scavenger, superoxide dismutase. The results suggest that inactivation of XO by H2O2 occurs by production of .OH following direct reduction of H2O2 by XO at the flavin site.
authors.affiliation: Webb-Waring Lung Institute, University of Colorado Health Sciences Center, Denver 80262.
authors.forename: L S
authors.initials: LS
authors.lastname: Terada
authors.forename: J A
authors.initials: JA
authors.lastname: Leff
authors.forename: D M
authors.initials: DM
authors.lastname: Guidot
authors.forename: I R
authors.initials: IR
authors.lastname: Willingham
authors.forename: J E
authors.initials: JE
authors.lastname: Repine
mesh_terms: D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005557:Formaldehyde / Q000378:metabolism; D016166:Free Radical Scavengers; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D006878:Hydroxides / Q000378:metabolism*; D017665:Hydroxyl Radical; D004369:Pentetic Acid / Q000494:pharmacology; D013379:Substrate Specificity; D013482:Superoxide Dismutase / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000378:metabolism*
chemical_list: D016166:Free Radical Scavengers; D006878:Hydroxides; D005557:Formaldehyde; D017665:Hydroxyl Radical; D004369:Pentetic Acid; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D004121:Dimethyl Sulfoxide