视网膜光感受器细胞外基质蛋白质SPACR的分离纯化  

Isolation and Purification of chick SPACR

作　　者：赵劲松[1] 黄岚峰[2] 王淑荣[1] 

机构地区：[1]吉林大学第二医院眼科医院眼底病一科,吉林长春130041 [2]吉林大学第二医院骨科,吉林长春130041

出　　处：《中国实验诊断学》2011年第11期1801-1803,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金资助项目(31071222);吉林省科技发展计划资助项目(项目编号20080738;200705242)

摘　　要：目的建立视网膜光感受器细胞外基质(Interphotoreceptor matrix,IPM)蛋白质sialoprotein associated with cones and rods(SPACR)分离和纯化方法。方法分离视网膜IPM,提取蛋白质组份,通过离子交换、凝胶过滤层析等技术分离纯化SPACR,并通过双向电泳及western blot技术验证。结果通过双向电泳技术及western blot结果显示分离纯化后仅有等电点在5-5.5,分子量约在150 kDa的单一条带和SPACR抗体反应,表明得到高纯度的SPACR。结论通过本文所述方法可以得到纯度较高的SPACR,为将来进一步对SPACR活性及结构研究奠定了基础。Objective To establish an improved method for isolation and purification of sialoprotein associated with cones and rods（SPACR）in interphotoreceptor matrix（IPM）.Methods Protein in PBS-insoluble IPM was extracted,SPACR was purified by DEAE-Sephacel and Sephacryl S-300 HR chromatography.Results The map of 2D gel electrophoresis showed that SPACR was in the section of pI 5-5.5 which the relative molecular weight was 150kDa.Purified SPACR was obtained.Conclusion A purification procedure was established in this study for SPACR with highly efficiency and simplicity by DEAE-Sephacel and Sephacryl S-300 HR chromatography.The present results would be valuable in analyzing biological activities and structural determination of SPACR.

关 键 词：视网膜 蛋白质提纯 双向电泳 

分 类 号：R774.1[医药卫生—眼科]