【摘要】 AID(APPintracellulardomain)是由淀粉样前体蛋白APP经γ分泌酶切割产生的胞内端,它可能参与细胞凋亡,基因转录等活动,并对阿尔茨海默氏症的形成有一定影响.为进一步研究AID的功能,在本实验中,我们用AID为诱饵蛋白,运用酵母双杂交系统筛选与其结合的蛋白,得到其中两个阳性克隆,一个为RPS21,另一个为FLJ20551.接着又在酵母体系用β半乳糖苷酶报告基因验证了它们的相互作用,并在哺乳动物细胞中用免疫共沉淀的方法再次验证了它们相互作用的特异性.这些结果表明RPS21和FLJ20551可能通过与AID相互作用而影响阿尔茨海默氏症的形成.更多还原

【Abstract】 Amyloid precursor protein (APP),a key protein in pathogenesis of Alzheimer’s Disease (AD),is a type Ⅰ transmembrane protein which can be cleaved by β- and γ-secretase to release the amyloidogenic β-amyloid peptides and the APP intracellular domain (AID).While Aβ has been widely believed to initiate pathogenic cascades culminating AD,the physiological functions of AID remain elusive.To study the function of AID,we attempted to identify its interacting proteins by Yeast Two-Hybrid screening using AID as the bait.Among many positive clones identified,two candidates are particularly of interest:one encodes ribosomal protein S21 (RPS21),and the other encodes a fragment of a novel protein FLJ20551.The interaction specificity with AID of both the fragment and the full-length of the preys was confirmed by using the β-galatosidase reporter.Furthermore,we demonstrated,in mammalian 293T cells,the successful expression of the clone encoding RPS21 and verified the interaction between RPS21 and AID by co-immunoprecipitation approach.Our study identified two new AID binding proteins,RPS21 and FLJ20551,and may assign novel roles for the two proteins in mediating APP/AID functions in physiological and pathological processes including AD pathogenesis.更多还原