Identification and purification of the aspartate/glutamate carrier from bovine heart mitochondria.
The aspartate/glutamate carrier from bovine heart mitochondria was solubilized with dodecyl-octaoxyethylene ether (C12E8) and purified by chromatography on hydroxyapatite and celite. On SDS gel electrophoresis, the purified aspartate/glutamate carrier consisted of a single protein band with an apparent Mr of 31,500. When reconstituted into liposomes the aspartate/glutamate carrier protein catalyzed an N-ethylmaleimide-sensitive aspartate/aspartate exchange. It was purified 620-fold with a recovery of 17.2% and a protein yield of 0.03% with respect to the mitochondrial extract. The properties of the reconstituted carrier, i.e. requirement for a counteranion, substrate specificity and inhibitor sensitivity, were similar to those of the aspartate/glutamate carrier as characterized in mitochondria.
Bisaccia
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Department of Pharmaco-Biology, University of Bari, Italy.
De Palma
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Palmieri
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D026922:Amino Acid Transport Systems, Acidic; D000818:Animals; D017920:Antiporters; D001224:Aspartic Acid / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002845:Chromatography; D003902:Detergents; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D005971:Glutamates / Q000378:metabolism*; D006882:Hydroxyapatites; D007700:Kinetics; D008081:Liposomes; D008929:Mitochondria, Heart / Q000378:metabolism*; D008970:Molecular Weight; D011510:Proteolipids
D026922:Amino Acid Transport Systems, Acidic; D017920:Antiporters; D002352:Carrier Proteins; D003902:Detergents; D005971:Glutamates; D006882:Hydroxyapatites; D008081:Liposomes; D011510:Proteolipids; C051046:aspartate-glutamate carrier; C020485:proteoliposomes; D001224:Aspartic Acid; D017886:Durapatite