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.