The extraction and reconstitution of the alpha-cyanocinnamate-sensitive pyruvate transporter from castor bean mitochondria.
The pyruvate carrier from castor bean mitochondria has been solubilized with Triton X-114 and partially purified using hydroxyapatite column chromatography. SDS-polyacrylamide gel electrophoresis of the hydroxyapatite-eluate showed that there were 6 major protein bands of Mr, 74kDa, 66kDa, 34kDa, 32kDa, 30kDa 12kDa. When the eluate was reconstituted into liposomes it was shown to catalyze a pyruvate exchange reaction which was sensitive to N-ethyl maleimide and a series of analogues of alpha-cyanocinnamate. The characteristics of this pyruvate exchange activity are similar to that found in intact mitochondria, and it is concluded that one or more proteins in the hydroxyapatite-eluate correspond to the pyruvate carrier.
D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D002367:Ricinus communis / Q000378:metabolism*; D002845:Chromatography / Q000379:methods; D002934:Cinnamates / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D026901:Membrane Transport Proteins; D008928:Mitochondria / Q000378:metabolism*; D027501:Monocarboxylic Acid Transporters; D010947:Plants, Toxic; D011773:Pyruvates / Q000378:metabolism*; D012278:Ricinus / Q000378:metabolism*; D012995:Solubility
D002352:Carrier Proteins; D002934:Cinnamates; D026901:Membrane Transport Proteins; D027501:Monocarboxylic Acid Transporters; D011773:Pyruvates; C018253:alpha-cyanocinnamate