Purification of the rat-liver mitochondrial dicarboxylate carrier by affinity chromatography on immobilized malate dehydrogenase.
The dicarboxylate carrier of rat-liver mitochondria, extracted by Triton X-100 and partially purified by hydroxylapatite chromatography, was retained by malate dehydrogenase immobilized on Sepharose gel, and eluted with 0.4 M NaCl. SDS-polyacrylamide gel electrophoresis of the eluate showed a predominant peptide band with an M(r) of 28,000. The purified protein, incorporated into liposomes, mediated a butylmalonate sensitive malonate(out)/malate(in) exchange that was inhibited by p-chloromercuriphenylsulfonate. Sulfate, malate and phosphate decreased the rate of exchange. The highly purified protein displayed all the properties of the dicarboxylate carrier. Moreover, the results suggest a possible functional interaction between mitochondrial carrier protein and malate dehydrogenase.
Lançar-Benba
J
J
University of Rouen, Faculty of Sciences, Laboratory of Intracellular Transports, URA CNRS No. 203, Mont-Saint-Aignan, France.
Foucher
B
B
Saint-Macary
M
M
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D029382:Dicarboxylic Acid Transporters; D004800:Enzymes, Immobilized; D008081:Liposomes; D008291:Malate Dehydrogenase / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D008314:Malonates / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism*; D017830:Octoxynol; D051381:Rats
D002352:Carrier Proteins; D029382:Dicarboxylic Acid Transporters; D004800:Enzymes, Immobilized; D008081:Liposomes; D008293:Malates; D008314:Malonates; D002731:4-Chloromercuribenzenesulfonate; C030298:malic acid; D017830:Octoxynol; C030290:malonic acid; D008291:Malate Dehydrogenase