PURIFICATION OF MYELIN CARBONIC ANHYDRASE

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Abstract
Abstract— A procedure has been developed for the purification of the membrane bound form of carbonic anhydrase from rat brain myelin. The procedure is rapid, requiring only two steps, and can be applied to small amounts of material. Conditions have been established whereby the enzyme can be almost quantitatively solubilized with up to a 60 fold increase in specific activity. Purification by affinity chromatography yields a preparation which is homogeneous by polyacrylamide gel electrophoresis. However, preliminary evidence suggests that activity may be reduced by the removal of lipids during chromatography and subsequent dialysis. The purified preparation is high in dicarboxylic and hydroxyl amino acids and contains only 1×2 cysteine residues. The reduction of cysteine appears to be essential for the full expression of enzymatic activity.