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- W2032033341 abstract "Despite the therapeutic potential of tempol (4-hydroxy-2,2,6,6-tetra-methyl-1-piperidinyloxy) and related nitroxides as antioxidants, their effects on peroxidase-mediated protein tyrosine nitration remain unexplored. This posttranslational protein modification is a biomarker of nitric oxide-derived oxidants, and, relevantly, it parallels tissue injury in animal models of inflammation and is attenuated by tempol treatment. Here, we examine tempol effects on ribonuclease (RNase) nitration mediated by myeloperoxidase (MPO), a mammalian enzyme that plays a central role in various inflammatory processes. Some experiments were also performed with horseradish peroxidase (HRP). We show that tempol efficiently inhibits peroxidase-mediated RNase nitration. For instance, 10 muM tempol was able to inhibit by 90% the yield of 290 muM 3-nitrotyrosine produced from 370 muM RNase. The effect of tempol was not completely catalytic because part of it was consumed by recombination with RNase-tyrosyl radicals. The second-order rate constant of the reaction of tempol with MPO compound I and II were determined by stopped-flow kinetics as 3.3 x 10(6) and 2.6 x 10(4) M(-1) s(-1), respectively (pH 7.4, 25 degrees C); the corresponding HRP constants were orders of magnitude smaller. Time-dependent hydrogen peroxide and nitrite consumption and oxygen production in the incubations were quantified experimentally and modeled by kinetic simulations. The results indicate that tempol inhibits peroxidase-mediated RNase nitration mainly because of its reaction with nitrogen dioxide to produce the oxammonium cation, which, in turn, recycles back to tempol by reacting with hydrogen peroxide and superoxide radical to produce oxygen and regenerate nitrite. The implications for nitroxide antioxidant mechanisms are discussed." @default.
- W2032033341 created "2016-06-24" @default.
- W2032033341 creator A5061850498 @default.
- W2032033341 creator A5064232275 @default.
- W2032033341 date "2008-06-17" @default.
- W2032033341 modified "2023-09-23" @default.
- W2032033341 title "Inhibition of myeloperoxidase-mediated protein nitration by tempol: Kinetics, mechanism, and implications" @default.
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- W2032033341 doi "https://doi.org/10.1073/pnas.0708211105" @default.
- W2032033341 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2448813" @default.
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