人源性表皮黑素细胞膜蛋白双向电泳技术初步建立及优化设计  

Preliminary establishment and optimization of two-dimensional electrophoresis for proteomic analysis of human epidermal melanocytes membrane

作　　者：李强[1] 焦彬 高天文[1] 于晓云[1] 阮禹松[3] 李春英[1] 王刚[1] 田艳丽[1] 刘玲[1] 

机构地区：[1]第四军医大学西京医院皮肤科,西安710032 [2]北京武警总队医院皮肤科,北京100027 [3]西安交通大学生命科学与技术学院,西安710049

出　　处：《中国麻风皮肤病杂志》2007年第2期118-121,共4页China Journal of Leprosy and Skin Diseases

摘　　要：目的:初步建立人源性表皮黑素细胞膜蛋白质组学研究中的双向电泳分离技术,优化膜蛋白提纯方法,以提高细胞膜蛋白的分辨率、敏感度和可重复性。方法:采用固相pH梯度(IPG)等电聚焦(IEF)为第一向、SDS-PAGE垂直电泳为第二向的双向电泳技术,对样品处理、水化等提取膜蛋白步骤进行了优化,与常规的提取膜蛋白的方法进行了比较。结果:经优化提纯方法所得样品经3次重复实验,获得了高分辨率的双向电泳图,可分离蛋白质斑点数达500多个,蛋白分布比较均匀、范围广,蛋白点没有脱尾现象,与常规提取膜蛋白的方法比较具有显著优势。结论:优化的蛋白提纯方法更适合细胞膜蛋白的蛋白组学研究。Objective： To establish two- dimensional electrophoris （2 - DE） for proteomic analysis of the human epidermal melanocytes membrane, and to optimize the purification method to promote resolution and reproducibility. Methods： A 2 - DE technique, including isoelectric focusing （IEF） with an immobilized pH gradient （IPG） and SDS- PAGE, was used. Sample preparation, rehydmtion, SDS - PAGE and other perfecting purification steps were established. The optimized purification method was compared with the routine method. Results： After the optimized purification was performed three times, a perfect 2- D map was gained with 500 protein spots, steadily distribution and wide range. The available character was that there were not protein spots tailing phenomena, which was much better than routine protein purification method. Conclusion： The optimized purification method is adapted to proteomic research on the cell membrane proteins.

关 键 词：蛋白质组学 双向电泳 膜蛋白 黑素细胞 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]