A simple purification of calmodulin by reversed phase chromatography with hydrophobic polymer 3520.
A new adsorption chromatography procedure for the purification of calmodulin from bovine brain was developed using polymeric adsorbent 3520. Calmodulin was first isolated by DEAE-Cellulose column chromatography and further purified to apparent homogeneity following elution with 50% ethanol from the adsorbent column. Polyacrylamide gel electrophoresis showed one band either in the presence of Ca2+ or EGTA. The polymeric adsorbent 3520 is a non-polar polymer lacking exchangeable groups. The selective adsorption of calmodulin is based on hydrophobic interaction within the matrix, and is Ca2+ independent. Neither high salt (0.5 M NaC1) nor EGTA (5 mM) was able to elute the CaM from the adsorption column whereas ethanol (50%) eluted it completely. This method is simple to use and it provides highly purified calmodulin with high yield.
D000818:Animals; D001923:Brain Chemistry; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000302:isolation & purification*; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D004533:Egtazic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D011137:Polystyrenes; D011145:Polyvinyls
D002147:Calmodulin; D011137:Polystyrenes; D011145:Polyvinyls; C051242:polymer 3520; D004533:Egtazic Acid; D002118:Calcium