INHIBITION OF CYTOCHROME-C-OXIDASE ACTIVITY BY 4-HYDROXYNONENAL (HNE)- ROLE OF HNE ADDUCT FORMATION WITH THE ENZYME SUBUNITS

The role of 4-hydroxynonenal (HNE), a major lipid peroxidation product in oxidative damage to mitochondrial cytochrome c oxidase (COX) was e xamined. Oxidative stress was induced in mitochondria isolated from li vers of male Sprague-Dawley rats by tert-butylhydroperoxide (t-BHP). C OX activity was inhibited, with a concommitant increase in endogenous HNE level in mitochondria. COX activity was also inhibited following i ncubation of mitochondria 50-150 mu M HNE. Blocking HNE degradation in tensified COX inhibition by HNE and by t-BHP-induced oxidative stress, the latter accompanied by a simultaneous increase in endogenous HNE p roduction. On the other hand, COX inhibition by HNE was markedly reduc ed by potentiating HNE degradation via enhancing conjugation of HNE wi th reduced glutathione (GSH). Incubation of purified COX with 10-400 m u M HNE resulted in HNE adduct formation with specific subunits of COX , correlated with inhibition of the enzyme activity. These data sugges t that HNE may inhibit mitochondrial COX by forming adducts with the e nzyme, and that this could be one mechanism underlying mitochondrial d amage caused by oxidative stress. The findings also illustrate a role for GSH in protecting mitochondria from the deleterious effects of HNE . (C) 1998 Elsevier Science B.V.