P57蛋白与HeLa细胞膜结合区的鉴定  

Identification of plasma membrane binding region of P57 with HeLa cell

作　　者：张祥[1] 金萌萌[2] 黄鹏[3] 张世谦[4] 张里程[3] 唐佩福[3] 

机构地区：[1]西安医学院附属第一医院骨外一科,陕西西安710077 [2]北京大学医学院第一临床医院内分泌科,北京100034 [3]中国人民解放军总医院,北京100853 [4]哈尔滨医科大学附属第一医院骨外一科,黑龙江哈尔滨150001

出　　处：《基础医学与临床》2011年第3期231-236,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(30672135);北京市自然科学基金(7072072)

摘　　要：目的研究肌动蛋白结合蛋白P57的性质、功能及其与细胞膜结合位点。方法用荧光显微镜观察野生P57蛋白及几种P57缺失突变体蛋白在细胞内的分布。利用细胞吹裂技术检测野生型P57及它的几种缺失突变体蛋白与细胞膜的结合。用差速离心技术分离亚细胞组分,用Western blot检测各组分中蛋白的含量。结果 P57蛋白的386-461片段具有与全长蛋白相类似的细胞分布及细胞膜结合能力,而1-299和1-432片段均不具有此能力。另外过量表达的386-461片段还可以竞争全长蛋白的膜结合位点。结论 P57是由其C末端区域(于386-461片段)负责蛋白与细胞膜的结合。Objective To study the property and function of P57 protein,and to identify the plasma membrane binding sites with HeLa cell.Methods The distribution of wild-type P57 and its mutants in cells were observed by fluorescence microscopy.The blowing crack technology was used to detect the binding between the wild-type P57 and its mutants with plasma membrane.Differential centrifugation technology was used to prepare subcellular components and the contents of P57 in subcellular components were detected by Western blot.Results It was found that the fragment 386-461 of P57 has a similar distribution in cells and similar plasma membrane binding abilility as the intact protein,but the fragment 1-299 or 1-432 hasn＇t.Overexpression of the fragment 386-461 can compete with P57 for membrane binding.Conclusion The C-terminal domain containing the fragment 386-461 is responsible for the binding of P57 to plasma membrane.

关 键 词：P57 细胞膜结合区域 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]