Identification and purification of the carnitine carrier from rat liver mitochondria.
The carnitine carrier from rat liver mitochondria, solubilized in Triton X-100 and partially purified on hydroxyapatite, was identified and completely purified by specific elution from celite in the presence of cardiolipin. On SDS-gel electrophoresis, the purified celite fraction consisted of a single band with an apparent Mr of 32,500. When reconstituted into liposomes the carnitine transport protein catalyzed an N-ethylmaleimide-sensitive carnitine/carnitine exchange. It was purified 970-fold with a recovery of 43% and a protein yield of 0.04% with respect to the mitochondrial extract. The properties of the reconstituted carrier, i.e., requirement for a countersubstrate, substrate specificity and inhibitor sensitivity, were similar to those of the carnitine transport system as characterized in intact mitochondria.
Indiveri
C
C
Department of Pharmaco-Biology, University of Bari, Italy.
Tonazzi
A
A
Palmieri
F
F
D000818:Animals; D001692:Biological Transport; D002308:Cardiolipins / Q000494:pharmacology; D002331:Carnitine / Q000378:metabolism*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008081:Liposomes / Q000302:isolation & purification; D008930:Mitochondria, Liver / Q000378:metabolism*; D008970:Molecular Weight; D011510:Proteolipids / Q000302:isolation & purification; D051381:Rats; D013379:Substrate Specificity
D002308:Cardiolipins; D008081:Liposomes; D011510:Proteolipids; C020485:proteoliposomes; D002331:Carnitine