The mitochondrial oxoglutarate carrier protein contains a disulfide bridge between intramembranous cysteines 221 and 224.
The oxoglutarate carrier (OGC) purified from bovine heart mitochondria was treated, both in its active and in its SDS-denatured state, with the fluorescent N-(1-pyrenyl)maleimide and other SH reagents before and after reduction with dithioerythritol or beta-mercaptoethanol. The number of SH groups per OGC polypeptide chain was found to be about 1 for the oxidized carrier and 3 for the reduced carrier. The bovine oxoglutarate carrier contains three cysteines: Cys-184, Cys-221 and Cys-224. Sequencing of BrCN cleavage products of oxoglutarate carrier showed that N-(1-pyrenyl)maleimide binds to only Cys-184 of the oxidized protein and also to Cys-221 and Cys-224 after reduction of the protein. These results show the presence of a disulfide bridge between the latter two cysteines of the purified carrier. The oxidized and the reduced forms of the oxoglutarate carrier exhibited different Vmax but virtually the same K(m) values for oxoglutarate.
Bisaccia
F
F
Department of Pharmaco-Biology, University of Bari, Italy.
Capobianco
L
L
Mazzeo
M
M
Palmieri
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D003545:Cysteine / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry*; D007425:Intracellular Membranes / Q000737:chemistry; D026901:Membrane Transport Proteins; D008929:Mitochondria, Heart / Q000737:chemistry*; D008969:Molecular Sequence Data
D002352:Carrier Proteins; D004220:Disulfides; D026901:Membrane Transport Proteins; C021723:oxoglutarate translocator; D003545:Cysteine