Identification and purification of the tricarboxylate carrier from rat liver mitochondria.
The tricarboxylate carrier from rat liver mitochondria was solubilized with Triton X-100 and purified by chromatography on hydroxyapatite and celite. SDS-gel electrophoresis of the purified fraction showed a single polypeptide band with an apparent Mr of 30,000. When reconstituted into liposomes, the tricarboxylate transport protein catalyzed a 1,2,3-benzenetricarboxylate-sensitive citrate/citrate exchange. We obtained a 1070-fold purification with respect to the mitochondrial extract, the recovery was 22% and the protein yield 0.02%. The properties of the reconstituted carrier, i.e., requirement for a counteranion, substrate specificity and inhibitor sensitivity, were similar to those of the tricarboxylate transport system as characterized in intact mitochondria.
Bisaccia
F
F
Department of Pharmaco-Biology, Laboratory of Biochemistry, University of Bari, Italy.
De Palma
A
A
Palmieri
F
F
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D002951:Citrates / Q000378:metabolism; D019343:Citric Acid; D004591:Electrophoresis, Polyacrylamide Gel; D008081:Liposomes; D008930:Mitochondria, Liver / Q000032:analysis*; D008970:Molecular Weight; D051381:Rats; D014233:Tricarboxylic Acids / Q000378:metabolism*
D002352:Carrier Proteins; D002951:Citrates; D008081:Liposomes; D014233:Tricarboxylic Acids; C020723:citrate-binding transport protein; D019343:Citric Acid