Maruyama K, Ebisawa K, Nonomura Y
Anal Biochem. 1985 Nov 15;151(1):1-6. doi: 10.1016/0003-2697(85)90043-0.
A vitamin D-dependent calcium-binding protein of 28,000 Mr was purified by a convenient method from the high-ammonium sulfate-saturated fraction (70 to 100% saturation) of bovine cerebellum and kidney. This method is based on the calcium-dependent DEAE-cellulose column chromatography that reflected the specific feature of this protein: different eluting salt concentrations in the presence or absence of Ca2+. The procedure of purification was easily performed by the detection of calcium-binding protein using 45Ca autoradiography. The purified calcium-binding protein showed the same molecular weight and calcium-dependent change of mobility in nondenaturing gel electrophoresis as vitamin D-dependent calcium-binding protein. The 28,000-Mr calcium-binding protein was induced in kidney by the injection of vitamin D in vitamin D-deficient rats.
通过一种简便方法，从牛小脑和肾脏的高硫酸铵饱和级分（70%至100%饱和度）中纯化出一种分子量为28,000的维生素D依赖性钙结合蛋白。该方法基于钙依赖性DEAE - 纤维素柱色谱，此方法反映了该蛋白的特定特性：在有或无Ca2+存在时洗脱盐浓度不同。通过使用45Ca放射自显影检测钙结合蛋白，很容易完成纯化过程。纯化后的钙结合蛋白在非变性凝胶电泳中显示出与维生素D依赖性钙结合蛋白相同的分子量和钙依赖性迁移变化。在维生素D缺乏的大鼠中注射维生素D后，肾脏中可诱导产生分子量为28,000的钙结合蛋白。
