Direct demonstration of a specific interaction between cyclophilin-D and the adenine nucleotide translocase confirms their role in the mitochondrial permeability transition.
A fusion protein between cyclophilin-D (CyP-D) and glutathione S-transferase (GST) was shown to bind to purified liver inner mitochondrial membranes (IMMs) in a cyclosporin A (CsA)-sensitive manner. Binding was enhanced by diamide treatment of the IMMs. Immobilized GST-CyP-D avidly bound a single 30 kDa protein present in Triton X-100-solubilized IMMs; immunoblotting showed this to be the adenine nucleotide translocase (ANT). Binding was prevented by pretreatment of the CyP-D with CsA, but not with cyclosporin H. Purified ANT also bound specifically to GST-CyP-D, but porin did not, even in the presence of ANT.
Woodfield
K
K
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
Rück
A
A
Brdiczka
D
D
Halestrap
A P
AP
D000818:Animals; D001278:Atractyloside / Q000031:analogs & derivatives / Q000494:pharmacology; D015153:Blotting, Western; D001865:Bongkrekic Acid / Q000494:pharmacology; D002846:Chromatography, Affinity; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins; D003958:Diamide / Q000494:pharmacology; D005982:Glutathione Transferase / Q000378:metabolism; D015151:Immunoblotting; D020104:Immunophilins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D007425:Intracellular Membranes / Q000187:drug effects; D008297:Male; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000226:Mitochondrial ADP, ATP Translocases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D017830:Octoxynol; D010539:Permeability; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D000081406:Peptidyl-Prolyl Isomerase F; D011993:Recombinant Fusion Proteins; D003958:Diamide; D001865:Bongkrekic Acid; D001278:Atractyloside; D017830:Octoxynol; D000226:Mitochondrial ADP, ATP Translocases; D005982:Glutathione Transferase; D021983:Cyclophilins; D020104:Immunophilins; C003853:carboxyatractyloside