An improved purification procedure for calpastatin, the inhibitor protein specific for the intracellular calcium-dependent proteinases, calpains.
An improved purification procedure for calpastatin, the inhibitor protein specific for the intracellular calcium-dependent proteinases, calpains.
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钙蛋白酶抑制剂的改进纯化程序，钙蛋白酶抑制剂是细胞内钙依赖性蛋白酶（钙蛋白酶）的特异性抑制剂蛋白。
DOI：
10.1080/00327488808062520
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Lane,RD
中科院分区：
文献类型：
--
作者：
Mellgren,RL;Nettey,MS;Mericle,MT;Renno,W;Lane,RD
钙依赖性细胞内蛋白酶（calpains）的特异性抑制蛋白（calpastatin）是这些酶的重要调节剂。在这篇文章中，我们描述了一个从一公斤牛心肌中纯化3至5毫克钙蛋白酶抑制蛋白的一天程序。这代表了对先前描述的方法的实质性改进，并且应该促进钙蛋白酶抑制蛋白结构和功能的未来研究。纯化中的一个关键的新步骤是Affi-Gel Blue柱上的染料基质色谱法。与以前的指示相反，通过新方法纯化的钙蛋白酶抑制素不含大量的碳水化合物。然而，在纯化的牛心肌钙蛋白酶抑制剂中共价结合的磷酸盐的存在被证实，并且在Bio-Gel A-1.5m色谱上证明了磷酸盐和钙蛋白酶抑制剂活性的共迁移。因此，钙蛋白酶抑制蛋白的功能可能是由磷酸化调节的。
The specific inhibitor protein (calpastatin) for the calcium-dependent intracellular proteinases (calpains) is an important regulator of these enzymes. In this communication we describe a one day procedure for purifying 3 to 5 mg of calpastatin from a kilogram of bovine myocardium. This represents a substantial improvement over previously described methods, and should facilitate future studies of calpastatin structure and function. A key, novel step in the purification was dye-matrix chromatography on an Affi-Gel Blue column. Contrary to previous indications, calpastatin purified by the new method did not contain significant amounts of carbohydrate. However, the presence of covalently bound phosphate in purified bovine myocardial calpastatin was confirmed and co-migration of phosphate and calpastatin activity was demonstrated on Bio-Gel A-1.5m chromatography. Thus, it is possible that calpastatin function is regulated by phosphorylation.