An ADP-sensitive cyclosporin-A-binding protein in rat liver mitochondria.
Mitochondria contain a structure which forms a large aqueous pore in the inner membrane after Ca2+ overload in the presence of Pi. In the present study, pore activation in liver mitochondria was monitored using the collapse of the inner membrane potential (delta psi). Ca(2+)-induced pore opening (delta psi collapse) was prevented by the immunosuppressant cyclosporin A, but cyclosporin A did not reverse pore opening (i.e. allow delta psi regeneration) unless ADP was also added. At concentrations that produced substantial pore blockade, [3H]cyclosporin partitioned more or less equally between membrane and soluble fractions, but the distribution was shifted slightly to the membranes in the presence of ADP. ADP also increased the binding of [3H]cyclosporin A to membranes washed free of soluble components. The indication that cyclosporin A inhibition of the pore is mediated by an ADP-sensitive membrane component was examined using a tritiated photoactivable derivative of cyclosporin A. ADP selectively increased covalent binding of this derivative to a membrane component. This component eluted from molecular-sizing columns as a 13-17-kDa-protein in the presence of 0.5% Chaps as detergent and migrated as a 10-kDa (approximately) protein in SDS/PAGE. These findings provide the first evidence that a protein of approximately 10 kDa may be part of the cyclosporin-A receptor of the Ca(2+)-activated pore. The possible implications of these findings are discussed.
Andreeva
L
L
Department of Biochemistry and Molecular Biology, University College London, England.
Crompton
M
M
D000244:Adenosine Diphosphate / Q000494:pharmacology*; D000345:Affinity Labels; D000591:Amino Acid Isomerases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002793:Cholic Acids; D016572:Cyclosporine / Q000378:metabolism / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism*; D019696:Peptidylprolyl Isomerase; D010743:Phospholipids / Q000378:metabolism; D010777:Photochemistry; D051381:Rats
D000345:Affinity Labels; D002352:Carrier Proteins; D002793:Cholic Acids; D008565:Membrane Proteins; D010743:Phospholipids; D000244:Adenosine Diphosphate; D016572:Cyclosporine; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; D002118:Calcium