Separation of calmodulin from calcium-activated protein kinase using calcium-dependent hydrophobic interaction chromatography.
Separation of calmodulin from calcium-activated protein kinase using calcium-dependent hydrophobic interaction chromatography.
复制标题
使用钙依赖性疏水相互作用色谱法从钙激活蛋白激酶中分离钙调蛋白。
DOI：
10.1016/0003-2697(88)90097-8
复制
发表时间：
1988
影响因子：
2.9
通讯作者：
M. Venis
中科院分区：
文献类型：
--
作者：
N. Battey;M. Venis
为了确定钙激活的蛋白激酶是否受钙调蛋白的调节，有必要将其与内源性钙调蛋白和非钙依赖的蛋白激酶活性分开。我们在这里描述了一种利用苯基琼脂糖凝胶上的钙离子依赖的疏水相互作用层析结合pH变化来实现这些目标的方法。这一过程是基于观察到的，当从苹果果膜中溶解的钙调蛋白在pH 7.0和8.5都以钙离子依赖的方式与苯基琼脂糖结合时，来自同一来源的钙激活的蛋白激酶仅在pH 7.5以上表现出钙离子依赖的相互作用。简要讨论了这一发现对调节这种钙激活的蛋白激酶的意义。
To determine whether a Ca2+-activated protein kinase is regulated by calmodulin, it is necessary to separate it from endogenous calmodulin and from protein kinase activity that is not calcium dependent. We describe here a procedure for achieving these goals using Ca2+-dependent hydrophobic interaction chromatography on phenyl Sepharose in combination with a pH change. The procedure is based on the observation that while calmodulin solubilized from apple fruit membranes binds to phenyl Sepharose in a Ca2+-dependent fashion at both pH 7.0 and 8.5, Ca2+-activated protein kinase from the same source only shows a Ca2+-dependent interaction above pH 7.5. The implications of this finding for the regulation of this Ca2+-activated protein kinase are briefly discussed.