视网膜光感受器细胞外基质蛋白质SPACR的分离纯化

摘要

0bjective To establish an in proved m ethod for isolation and purification of siabprotein associated w ith cones and rods(SPACR )in interohotoreceptar m ateix ( IPM ) .M eihods Protein in PBS-insoluble IPM was extracted ,SPACR was purified by DEAE-Sephaceland SephacrylS-300 HR chrom atography .R esults Themap of 2D gel electtophoresis showed thatSPACR was in the section of pI 5-5 .5 which the relative molecular weight was 150kD a. Purified SPACR was obtained. C onclision A purification procedure was established in this study for SPACR with highly efficiency and sinplicity by DEAE-Sephacel and Sephacryl S-300 HR chrom atogiaphy .The present results would be valuable in analyzing biobgical activities and structural detem ination of SPACR .%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.