Peroxidative damage to cardiac mitochondria: identification and purification of modified adenine nucleotide translocase.
Rat myocardial membranes exposed to free radical-generating systems exhibit both lipid peroxidation and protein alterations. The most sensitive protein, a 28-kDa polypeptide, was previously shown to increase slightly in apparent molecular weight before disappearing completely from the protein profile [N. L. Parinandi, C. W. Zwizinski, and H. H. O. Schmid (1991) Arch. Biochem. Biophys. 289, 118-123]. We now report that isolated cardiac mitochondria contain a 28-kDa protein which responds in the same manner to treatment with Cu2+/t-butylhydroperoxide. The protein exhibits several characteristic properties of the mitochondrial adenine nucleotide translocase. This assignment is supported by the finding that carboxyatractyloside, a specific inhibitor of the adenine nucleotide translocase, can prevent the oxidant-induced changes in the 28-kDa protein. Efficient purification schemes for the isolation of milligram quantities of unmodified and oxidatively altered adenine nucleotide translocase from rat heart mitochondria are described.
Zwizinski
C W
CW
Hormel Institute, University of Minnesota, Austin 55912.
Schmid
H H
HH
D000818:Animals; D001278:Atractyloside / Q000031:analogs & derivatives / Q000494:pharmacology; D002845:Chromatography; D003300:Copper / Q000494:pharmacology*; D005609:Free Radicals; D015227:Lipid Peroxidation; D008929:Mitochondria, Heart / Q000187:drug effects / Q000201:enzymology*; D000226:Mitochondrial ADP, ATP Translocases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D010545:Peroxides / Q000494:pharmacology*; D051381:Rats; D020122:tert-Butylhydroperoxide
D005609:Free Radicals; D010545:Peroxides; D001278:Atractyloside; D003300:Copper; D000226:Mitochondrial ADP, ATP Translocases; D020122:tert-Butylhydroperoxide; C003853:carboxyatractyloside