Separation of membrane protein-sodium dodecyl sulphate complexes by high-performance liquid chromatography on hydroxyapatite

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Abstract The analytical conditions for the high-performance liquid chromatography of protein-sodium dodecyl sulphate complexes on ceramic hydroxyapatite were optimized for the purification of membrane proteins and the microanalysis of membrane protein mixtures. About 12 mg of erythrocyte membrane protein could be separated by ceramic hydroxyapatite chromatography at one time and about 0.8 mg of purified anion carrier protein was obtained on one-step purification. Erythrocyte membrane proteins could be analysed with buffers not containing dithiothreitol and detection at 230 nm. A two-step method, i.e., combined diethylaminoethyl and hydroxyapatite chromatography, made it possible to separate many membrane proteins.

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