BIOCHEMICAL STUDIES OF CALRETININ

这项工作的目的是确定的生化功能， 钙视网膜蛋白。初步结果描述了一些基本的 钙视网膜蛋白的特性，并将作为进一步的基础， 问题研究这些包括以下内容：（1）钙视网膜蛋白被确定在 用放射免疫分析法和 免疫印迹。钙使钙视网膜蛋白的量增加 与突触膜部分相关，并降低了 细胞质中钙视黄蛋白相对于EGTA存在下发现的钙视黄蛋白;（2） 有几种方法被用来寻找calretinin靶蛋白。 发现碘化钙视网膜蛋白与几种亚细胞中的蛋白质结合， 一维SDS凝胶硝酸纤维素印迹上的组分;（3）A 开发了纯化程序以获得大量的 来自质粒的同源重组钙视网膜蛋白。无差异 在天然和重组的理化性质中发现 钙视网膜蛋白（分子量，等电点，钙结合动力学 和在离子交换上的行为：凝胶过滤和疏水柱）; (4)一系列实验使用特定的树脂来研究疏水性 calretinin和树脂的性质。部分钙依赖 对某些树脂（辛基琼脂糖）观察到的钙视黄蛋白结合是 取决于离子强度和pH。

This work was aimed towards determining the biochemical function of calretinin. The initial results describe some of the basic characteristics of calretinin and will serve as the basis of further studies. These include the following: (1) Calretinin was identified in membrane fractions of rat cerebellum by both radioimmunoassay and immunoblot. Calcium produced an increase in the amount of calretinin associated with the synaptic membrane fraction and decreased the calretinin in cytosol relative to that found in the presence of EGTA; (2) Several approaches were used to search for calretinin target proteins. Iodinated calretinin was found to bind to proteins in several subcellular fractions on nitrocellulose blots of one dimensional SDS gels; (3) A purification procedure was developed for obtaining large amounts of homogeneous recombinant calretinin from plasmids. No differences were found in the physicochemical properties of native and recombinant calretinin (molecular weight, isoelectric point, calcium binding kinetics and behavior on ion exchange: gel filtration and hydrophobic columns); (4) A series of experiments used specific resins to study the hydrophobic properties of calretinin and the resins. A partial calcium dependence of calretinin binding observed for some resins (octylsepharose) was dependent on ionic strength and pH.