Mitochondrial stress protein recognition of inactivated dehydrogenases during mammalian cell death.
The mammalian renal toxicant tetrafluoroethylcysteine (TFEC) is metabolized to a reactive intermediate that covalently modifies the lysine residues of a select group of mitochondrial proteins, forming difluorothioamidyl lysine protein adducts. Cellular damage is initiated by this process and cell death ensues. NH2-terminal sequence analysis of purified mitochondrial proteins containing difluorothioamidyl lysine adducts identified the lipoamide succinyltransferase and dihydrolipoamide dehydrogenase subunits of the alpha-ketoglutarate dehydrogenase complex (alphaKGDH), a key regulatory component of oxidative metabolism, as targets for TFEC action. Adduct formation resulted in marked inhibition of alphaKGDH enzymatic activity, whereas the related pyruvate dehydrogenase complex was unmodified by TFEC and its activity was not inhibited in vivo. Covalent modification of alphaKGDH subunits also resulted in interactions with mitochondrial chaperonin HSP60 in vivo and with HSP60 and mitochondrial HSP70 in vitro. These observations confirm the role of mammalian stress proteins in the recognition of abnormal proteins and provide supporting evidence for reactive metabolite-induced cell death by modification of critical protein targets.
Bruschi
S A
SA
Department of Medicinal Chemistry, University of Washington, Seattle, WA 98195-7610, USA. sambru@u.washington.edu
Lindsay
J G
JG
Crabb
J W
JW
D000217:Acyltransferases / Q000378:metabolism; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D008058:Dihydrolipoamide Dehydrogenase / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006845:Hydrocarbons, Fluorinated / Q000378:metabolism / Q000494:pharmacology*; D007655:Ketoglutarate Dehydrogenase Complex / Q000235:genetics / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism / Q000473:pathology*; D008928:Mitochondria / Q000378:metabolism* / Q000473:pathology*; D051381:Rats; D005466:Fluorocarbons
D000217:Acyltransferases; D003545:Cysteine; D008058:Dihydrolipoamide Dehydrogenase; D006360:Heat-Shock Proteins; D006845:Hydrocarbons, Fluorinated; D007655:Ketoglutarate Dehydrogenase Complex; C064116:S-(1,1,2,2-tetrafluoroethyl)cysteine; D005466:Fluorocarbons; C021515:dihydrolipoamide succinyltransferase