Purification of rat liver mitochondrial anion carriers by triazine dye affinity chromatography.
Mitochondria were prepared from rat liver, solubilized with Triton X-100 and submitted to chromatography on dry hydroxylapatite. The pass-through from the column, containing a mixture of mitochondrial carriers, was processed further by triazine dye affinity chromatography and optimal conditions for binding and elution of mitochondrial carriers were established. Activities of the mono-, di-, tricarboxylate and 2-oxoglutarate carriers were measured after reconstitution of various chromatographic fractions into asolectin vesicles, and a good resolution of different carriers was obtained. Thus the triazine dye affinity chromatography appears to be a potent new tool for purification of mitochondrial anion carriers.
Szewczyk
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Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warszawa, Poland.
Poddana
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Nałecz
M J
MJ
D000818:Animals; D027321:Anion Transport Proteins; D002264:Carboxylic Acids / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002845:Chromatography; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D006882:Hydroxyapatites; D008081:Liposomes / Q000378:metabolism; D008930:Mitochondria, Liver / Q000378:metabolism*; D011510:Proteolipids / Q000378:metabolism; D051381:Rats; D014227:Triazines
D027321:Anion Transport Proteins; D002264:Carboxylic Acids; D002352:Carrier Proteins; D004396:Coloring Agents; D006882:Hydroxyapatites; D008081:Liposomes; D011510:Proteolipids; D014227:Triazines; C020485:proteoliposomes; C012842:Cibacron Blue F 3GA; D017886:Durapatite