Rapid purification of calmodulin and S-100 protein by affinity chromatography with melittin immobilized to sepharose.
Melittin-Sepharose was prepared for Ca2+-dependent affinity chromatography of calmodulin and S-100 protein. This matrix exhibits extremely high capacity (approximately 10 mg calmodulin/ml gel), low nonspecific binding, and excellent recovery (greater than 90%) under optimal conditions. Recovery of calmodulin from melittin-Sepharose was related to the degree of saturation of column capacity with lower yields when only partial saturation was achieved. Large-scale, simultaneous purification of calmodulin and S-100 protein from brain was carried out using selective adsorption to organomercurial agarose followed by melittin-Sepharose chromatography; yields were 250-300 mg of calmodulin and 200-300 mg of S-100 per kg tissue. Calmodulin also was purified in a single step from bovine testis supernatant using melittin-Sepharose in yields comparable to those from brain.
D000818:Animals; D001514:Bee Venoms / Q000494:pharmacology*; D001923:Brain Chemistry; D002147:Calmodulin / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D008297:Male; D008555:Melitten / Q000494:pharmacology*; D009418:S100 Proteins / Q000302:isolation & purification*; D012685:Sepharose / Q000494:pharmacology; D013737:Testis / Q000032:analysis
D001514:Bee Venoms; D002147:Calmodulin; D009418:S100 Proteins; D008555:Melitten; D012685:Sepharose