Mechanism in the reaction of cytochrome c oxidase with organic hydroperoxides: an ESR spin-trapping investigation

The Biochemical Journal
Yeong-Renn Chen, R P Mason

Abstract

Organic hydroperoxides are of great utility in probing the reaction mechanism and the toxicological consequences of lipid peroxidation. In the present study, ESR spin-trapping was employed to investigate the peroxidation of mitochondrial cytochrome c oxidase (CcO) with t-butyl hydroperoxide (t-BuOOH) and cumene hydroperoxide (CumOOH). The spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was used to detect the radical species formed from the reaction of CcO with t-BuOOH. The presence of t-BuOOH-derived alkoxyl radical (t-BuO*) as the primary radical indicates reductive scission of the O-O bond by CcO. The ESR signal of DMPO/*Ot-Bu can be partially abolished by cyanide, implying that the reductive cleavage involved the haem a(3)Cu(B) binuclear site of CcO. A nitroso spin trap, 2-methyl-2-nitrosopropane (MNP), was used to detect and identify radical species from the reaction of CcO with CumOOH. In addition to the t-BuOOH-derived methyl, hydroxylmethyl and tertiary carbon-centred radicals, a protein-derived radical was detected. The intensity of the ESR signal from the protein radical increased with the CumOOH concentration at low CumOOH/CcO ratios, with maximal intensity at a ratio of 100 mol of CumOOH/mol of CcO. The immobilized...Continue Reading

Citations

Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Ben-Zhan ZhuBalz Frei
Oct 31, 2007·Proceedings of the National Academy of Sciences of the United States of America·Ben-Zhan ZhuGui-Bin Jiang
Apr 10, 2013·Biochimica Et Biophysica Acta·Clare L Hawkins, Michael J Davies
Feb 22, 2005·Biochimica Et Biophysica Acta·Dimitri A Svistunenko
Apr 15, 2004·Free Radical Biology & Medicine·Michael J Davies, Clare L Hawkins
Apr 12, 2005·Archives of Biochemistry and Biophysics·Honglian ShiKe Jian Liu
Jun 1, 2005·Journal of Magnetic Resonance·Chunqi ChangFrancis H Y Chan
Jun 21, 2005·Archives of Biochemistry and Biophysics·Yeong-Renn ChenJay L Zweier

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