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利用DEAE-Sephadex和Phenyl-Sepharose层析,同时从猪脑中分离纯化了两种钙结合蛋白,即S-100和CaM。分子量为10,500的钙结合蛋白对牛脑PDE无激活作用;与CaM抗体无交叉反应;在SDS-PAGE中,迁移速度不受Ca~(2+)的影响;而在碱性甘油PAGE中,迁移速度在有Ca~(2+)时比无Ca~(2+)时慢,并且出现多区带;在250、260、266、270、279和284nm处具有六个吸收峰。以上特征及其氨基酸组成分析表明,此蛋白为S—100。另一种钙结合蛋白分子量为18,000;对牛脑PDE的激活作用和电泳行为与标准CaM相同;与CaM抗体呈阳性反应。
Using DEAE-Sephadex and Phenyl-Sepharose chromatography, two calcium-binding proteins, S-100 and CaM, were isolated and purified simultaneously from porcine brain. Calcium-binding protein with molecular weight of 10,500 showed no activation effect on PDE of bovine brain, no cross-reaction with CaM antibody, migration speed was not affected by Ca 2+ in SDS-PAGE, and migration in alkaline glycerol PAGE The rate was slower with Ca 2+ than with Ca 2+ and appeared in multiple bands; there were six absorption peaks at 250, 260, 266, 270, 279 and 284 nm. The above characteristics and amino acid composition analysis showed that the protein was S-100. Another calcium-binding protein has a molecular weight of 18,000; the activation and electrophoresis of PDE in bovine brain are the same as that of standard CaM; positive reaction with CaM antibody.