Rapid purification by affinity chromatography of rat brain pyridoxal kinase and pyridoxamine-5-phosphate oxidase |
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Authors: | C D Cash M Maitre J F Rumigny P Mandel |
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Institution: | 1. Centre de Neurochimie, Faculté de Médecine, 11 rue Humann, 67085 Strasbourg Cedex, France;2. Inserm U 44, Faculté de Médecine, 11 rue Humann, 67085 Strasbourg Cedex, France |
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Abstract: | Rat brain pyridoxal kinase and pyridoxamine 5′ phosphate oxidase have been purified to electrophoretic homogeneity using pyridoxyl Sepharose and phosphopyridoxyl Sepharose columns as the first stages in the purification procedures. These affinity supports were synthesized by two simple steps consisting of reacting pyridoxal and pyridoxal 5′ phosphate with commercially available ω-aminohexyl Sepharose and subsequent reduction of the resultant Schiff's bases with sodium borohydride. This method allows total purification of both enzymes from the same tissue source in 4–5 days. |
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