Moore P B
Biochem J. 1986 Aug 15;238(1):49-54. doi: 10.1042/bj2380049.
A set of four proteins, termed calcimedins, are isolatable from smooth, cardiac and skeletal muscle by using a fluphenazine-Sepharose affinity column. The calcimedins show apparent Mr values of 67,000, 35,000, 33,000 and 30,000 by SDS/polyacrylamide-gel electrophoresis. The 67,000-Mr calcimedin (67 kDa calcimedin) has now been purified to homogeneity by using DEAE-cellulose chromatography followed by Ca2+-dependent binding to phenyl-Sepharose. The amino acid analysis of the 67 kDa calcimedin shows this protein does not contain trimethyl-lysine but does contain 2 mol of tryptophan/mol of protein. The 67 kDa calcimedin shows positive ellipticity in the near-u.v. range with c.d. Ca2+-binding studies indicate one high-affinity Ca2+-binding site with Kd 0.4 microM. The data show that the 67 kDa calcimedin is distinct from other Ca2+-binding proteins described to date.
通过使用氟奋乃静 - 琼脂糖亲和柱,可从平滑肌、心肌和骨骼肌中分离出一组四种蛋白质,称为钙调素。通过SDS /聚丙烯酰胺凝胶电泳,钙调素的表观分子量值分别为67,000、35,000、33,000和30,000。现在,通过使用DEAE - 纤维素色谱法,然后进行依赖于Ca2 +的苯基 - 琼脂糖结合,已将67,000分子量的钙调素(67 kDa钙调素)纯化至同质。67 kDa钙调素的氨基酸分析表明,该蛋白质不含三甲基赖氨酸,但每摩尔蛋白质含有2摩尔色氨酸。67 kDa钙调素在近紫外范围内具有正椭圆率,通过圆二色性检测。Ca2 +结合研究表明存在一个高亲和力的Ca2 +结合位点,解离常数Kd为0.4 microM。数据表明,67 kDa钙调素与迄今为止描述的其他Ca2 +结合蛋白不同。