早老素1与热休克蛋白70同源蛋白羧基端相互作用蛋白  

Study on the interaction between presenilin 1 and carboxyl terminus of Hsc70 interacting protein

作　　者：刘润忠[1] 易雪莲[1] 霍淑芳[1] 扬悦[1] 侯海波[1] 张云武[1] 洪水根[1] 许华曦[1] 

机构地区：[1]福建省厦门市,厦门大学生命科学学院生物医学研究院分子与细胞神经科学实验室,361005

出　　处：《中华老年医学杂志》2008年第10期766-769,共4页Chinese Journal of Geriatrics

基　　金：国家自然科学基金（30572077）;细胞生物学与肿瘤细胞工程教育部重点实验室基金（2005111）

摘　　要：目的探讨早老素（PS1）蛋白突变体在阿尔茨海默病（AD）发生中的作用和PS1蛋白的正常生理功能及分子伴侣蛋白热休克蛋白70同源蛋白羧基端相互作用蛋白（CHIP）的相互作用。方法采用酵母双杂交系统筛选与PS1蛋白相互作用的蛋白。在构建pGBKT7-PS1-C203诱饵质粒和含全长CHIP的pACT2-CHIP质粒表达载体后，用β-半乳糖苷酶活性测定法检测两者的相互作用；然后转染哺乳动物细胞293T，用Co-IP和Western blot法测试其相互作用。结果获得了一个能与PS1蛋白相互作用的蛋白，即CHIP，并通过β-半乳糖苷酶活性测试、免疫共沉淀进一步证实了PS1和CHIP相互作用的特异性。结论CHIP既可以和分子伴侣蛋白相互作用，本身又具有泛素连接酶活性，可调控蛋白的折叠和降解，PS1与CHIP相互作用的证实，有助于阐明机体泛素-蛋白酶体系统对PS1的调控和进一步阐明AD的病理机制。Objective To understand the pathological and physiological roles of Presenilin 1 （PS1） in Alzheimer＇ s disease （AD） recurrence, and the interaction between PS1 and carboxyl terminus of Hse70 interacting protein （CHIP）. Methods The yeast two hybrid system was applied to identify a novel PS1 interacting protein as CHIP. After pGBKT7-PS1-C203 bait plasmid and full fragement CHIP of pACT2-CHIP expression vector were constructed, the interaction between PSI and CHIP was tested by β-galactosidase assay, pGBKT7 PS1 C203 was co-transfected with pACT2 -CHIP into 293T cells and the interaction between PS1 and CHIP was tested by coimmunoprecipitation and Western blot. Results Specificity of the interaction between PSI and CHIP was identified by β-galactosidase assay and co-immunoprecipitation. Conclusions CHIP is able to modulate chaperone functions and the pathway of protein ubiquitination/degradation. CHIP may regulate a proper assembly of the γ-secretase complex through its interaction with PS1, which is helpful to elucidate the mechanism of AD pathology.

关 键 词：阿尔茨海默病 早老素1 热休克蛋白质类 

分 类 号：R686[医药卫生—骨科学]