豚鼠心肌组织Cav1.2钙通道蛋白的提取与纯化  

Purification of cardiac Cav1. 2 channel protein from guinea pig heart

作　　者：封瑞[1] 于丽凤[1] 胡慧媛[1] 郭凤[1] 龟山正树[2] 郝丽英[1] 

机构地区：[1]中国医科大学药学院药物毒理学教研室,辽宁沈阳110001 [2]鹿儿岛大学医学部

出　　处：《中国药理学通报》2013年第11期1610-1613,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31071004;30870907;81100108)

摘　　要：目的探寻从豚鼠心肌组织中提取纯化全长的心肌Cav1.2钙通道蛋白的实验方法。方法采用组织匀浆、超高速离心的方法获得溶解状态的Cav1.2钙通道蛋白,使用亲和层析的方法对钙通道蛋白进行进一步的纯化,再应用Western blot技术使用不同的Cav1.2钙通道抗体对所制备的心肌Cav1.2钙通道蛋白进行鉴定。结果比较CHAPS和Triton X-100的增溶效果,发现去污剂CHAPS能够更好地促进钙通道蛋白的溶解,且浓度为2%CHAPS是能够使钙通道蛋白溶解且不损害钙通道结构的最适浓度;同时,蛋白电泳和免疫印迹结果显示,提取制备所得的蛋白为心肌Cav1.2钙通道蛋白。结论亲和层析方法提取了纯度较高的全长的心肌Cav1.2钙通道蛋白,为系统研究心肌Cav1.2钙通道奠定了坚实的基础。Aim To explore the novel methods for purification of cardiac Cavl. 2 channel protein from guinea pig heart. Methods The soluble Cavl. 2 channel protein was purified by tissue homogenate and uhracentrifugation methods, and affinity chromatography was used for further purification of Carl. 2 channel protein. Moreover, the Carl. 2 channel protein was detected by Western blot. Results Comparison of the soluble effect between CHAPS and Triton X-100 showed that 2% CHAPS was more suitable for dissolving the Cavl. 2 channel protein without breaking up the protein structure. Furthermore, the Cav1. 2 channel protein was identified by SDS-PAGE electrophoresis and Western blot. Conclusions The full length Cavl. 2 channel protein was purified by affinity chromatography method, and the purity was high.

关 键 词：Cav1 2钙通道 蛋白 心肌 豚鼠 亲和层析 表面活性剂 

分 类 号：R-332[医药卫生] R322.11