Mellgren R L, Nettey M S, Mericle M T, Renno W, Lane R D
Department of Pharmacology and Therapeutics, Medical College of Ohio, Toledo 43699.
Prep Biochem. 1988;18(2):183-97. doi: 10.1080/00327488808062520.
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.
钙依赖性细胞内蛋白酶（钙蛋白酶）的特异性抑制蛋白（钙蛋白酶抑制蛋白）是这些酶的重要调节因子。在本通讯中，我们描述了一种从一千克牛心肌中纯化3至5毫克钙蛋白酶抑制蛋白的一日程序。这比先前描述的方法有了实质性改进，应有助于未来对钙蛋白酶抑制蛋白结构和功能的研究。纯化过程中的一个关键新步骤是在Affi-Gel Blue柱上进行染料基质色谱法。与先前的迹象相反，通过新方法纯化的钙蛋白酶抑制蛋白不含大量碳水化合物。然而，证实了纯化的牛心肌钙蛋白酶抑制蛋白中存在共价结合的磷酸盐，并在Bio-Gel A-1.5m色谱法上证明了磷酸盐与钙蛋白酶抑制蛋白活性的共同迁移。因此，钙蛋白酶抑制蛋白的功能可能受磷酸化调节。
