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Free Radic Biol Med. 1993 May;14(5):559-65.

Alkoxyl and methyl radical formation during cleavage of tert-butyl hydroperoxide by a mitochondrial membrane-bound, redox active copper pool: an EPR study.

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1
Dept. Bioquimica de la Nutrición, Instituto Superior de Investigaciones Biológicas, San Miguel de Tucumàn, Argentina.

Abstract

The cleavage of tert-butyl hydroperoxide by submitochondrial particles yielded two distinctive radicals, alkoxyl and methyl radicals, detected by the electron paramagnetic resonance technique in conjunction with the spin trap 5,5'-dimethyl-1-pyrrolyne-N-oxide. Free radical formation was partly sensitive to bathocuproine disulfonate and was augmented upon supplementation of the mitochondrial membranes with copper, thus suggesting that a redox active copper pool in mitochondrial membranes participated actively in the cleavage of the O-O bond of tert-butyl hydroperoxide. This view was experimentally substantiated by the following: first, observation of the maximal EPR signal intensity required the presence of exogenous electron donors, such as succinate or reduced nicotinamide adenine dinucleotide (NADH). Second, copper reduction was accomplished partly by a superoxide radical-dependent mechanism as indicated by the sensitivity of the electron paramagnetic resonance (EPR) signal to superoxide dismutase. Third, the enhancing effect of the respiratory chain inhibitors, antimycin A and rotenone, on free radical yield assessed in the presence of superoxide dismutase pointed to the occurrence of two potential loci in the respiratory chain involved in direct electron transfer to membrane-bound copper: one located between NADH and the rotenone-sensitive site and another, quantitatively less important, between the rotenone- and antimycin-sensitive sites. These results support the notion that a redox pool of copper tightly bound to the mitochondrial membrane contributes significantly to the reductive cleavage of organic peroxides associated with free radical production.

PMID:
8394271
[Indexed for MEDLINE]
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