Identification and purification of the ornithine/citrulline carrier from rat liver mitochondria.
The ornithine/citrulline carrier from rat liver mitochondria, solubilized with Triton X-100 and partially purified on hydroxyapatite, was identified and completely purified by PD-10, DEAE-Sephacel and celite chromatography. On SDS/polyacrylamide gel electrophoresis, the purified ornithine/citrulline carrier consisted of a single protein band with an apparent molecular mass of 33.5 kDa. When reconstituted into liposomes the ornithine carrier protein catalyzed an active mersalyl sensitive ornithine/ornithine exchange. It was purified 438-fold with a recovery of 38% and a protein yield of 0.09% with respect to the extract derived from mitoplasts. The purified and reconstituted protein did not catalyze a significant unidirectional transport of ornithine. Citrulline was found to be the best countersubstrate for the transport of ornithine, followed by lysine and arginine. The exchange activity was inhibited by several sulphydryl reagents.
Indiveri
C
C
Department of Pharmaco-Biology, University of Bari, Italy.
Tonazzi
A
A
Palmieri
F
F
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002308:Cardiolipins / Q000737:chemistry; D002956:Citrulline / Q000378:metabolism*; D003902:Detergents; D007425:Intracellular Membranes / Q000737:chemistry; D008081:Liposomes; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008930:Mitochondria, Liver / Q000737:chemistry*; D008970:Molecular Weight; D009952:Ornithine / Q000378:metabolism*; D051381:Rats
D002308:Cardiolipins; D003902:Detergents; D008081:Liposomes; D008565:Membrane Proteins; D002956:Citrulline; D009952:Ornithine