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  • Title: Hydroxyl radical production from hydrogen peroxide and enzymatically generated paraquat radicals: catalytic requirements and oxygen dependence.
    Author: Winterbourn CC, Sutton HC.
    Journal: Arch Biochem Biophys; 1984 Nov 15; 235(1):116-26. PubMed ID: 6093705.
    Abstract:
    The ability of paraquat radicals (PQ+.) generated by xanthine oxidase and glutathione reductase to give H2O2-dependent hydroxyl radical production was investigated. Under anaerobic conditions, paraquat radicals from each source caused chain oxidation of formate to CO2, and oxidation of deoxyribose to thiobarbituric acid-reactive products that was inhibited by hydroxyl radical scavengers. This is in accordance with the following mechanism derived for radicals generated by gamma-irradiation [H. C. Sutton and C. C. Winterbourn (1984) Arch. Biochem. Biophys. 235, 106-115] PQ+. + Fe3+ (chelate)----Fe2+ (chelate) + PQ++ H2O2 + Fe2+ (chelate)----Fe3+ (chelate) + OH- + OH.. Iron-(EDTA) and iron-(diethylenetriaminepentaacetic acid) (DTPA) were good catalysts of the reaction; iron complexed with desferrioxamine or transferrin was not. Extremely low concentrations of iron (0.03 microM) gave near-maximum yields of hydroxyl radicals. In the absence of added chelator, no formate oxidation occurred. Paraquat radicals generated from xanthine oxidase (but not by the other methods) caused H2O2-dependent deoxyribose oxidation. However, inhibition by scavengers was much less than expected for a reaction of hydroxyl radicals, and this deoxyribose oxidation with xanthine oxidase does not appear to be mediated by free hydroxyl radicals. With O2 present, no hydroxyl radical production from H2O2 and paraquat radicals generated by radiation was detected. However, with paraquat radicals continuously generated by either enzyme, oxidation of both formate and deoxyribose was measured. Product yields decreased with increasing O2 concentration and increased with increasing iron(DTPA). These results imply a major difference in reactivity between free and enzymatically generated paraquat radicals, and suggest that the latter could react as an enzyme-paraquat radical complex, for which the relative rate of reaction with Fe3+ (chelate) compared with O2 is greater than is the case with free paraquat radicals.
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