FasL相关蛋白SH3P12影响FasL细胞膜表达效应研究  

FasL associated factor SH3P12 and its potential role in the regulation of FasL expression

作　　者：钱景[1] 彭慧琴[1] 周倩[2] 王巧玲[2] 陈浩浩[2] 严杰[1] 

机构地区：[1]浙江大学医学院病原生物学教研室,杭州310058 [2]浙江大学医学院基础医学系2002级,杭州310058

出　　处：《中华微生物学和免疫学杂志》2006年第11期1013-1016,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的研究FasL相关蛋白SH3P12对FasL表达的影响。方法构建SH3P12真核细胞表达载体,并与FasL共同短暂表达于人胚肾细胞293T细胞。以免疫共沉淀和免疫印迹法确定目的蛋白之间的相互作用,以免疫荧光染色结合激光共聚焦显微镜观察SH3P12与FasL在细胞内表达。结果在转染的293T细胞中,SH3P12与FasL可形成免疫共沉淀的蛋白-蛋白复合物。荧光蛋白标记结合显微镜观察提示在无SH3P12存在时,FasL主要分布于293T细胞膜,并伴随部分蛋白表达在细胞内高尔基体或内含体样颗粒中；在有SH3P12存在时,FasL与SH3P12形成复合体,共同定位于细胞膜,此时细胞内表达FasL量明显降低。结论SH3P12为FasL细胞内结合蛋白,其生物学活性可能为稳定FasL在细胞膜上的表达。Objective To investigate the role of a putative FasL interacting protein SH3P12 in the context of FasL expression and function. Methods A myc. his tagged mammalian expression vector encoding the full length of SH3P12 fragment was constructed by the standard RT-PCR based cloning procedure. Calcium phosphate precipitation method was used to transfect 293T cells for transient expression. Protein-protein interacting was analyzed by an anti-FasL or anti-myc co-immunoprecipitation followed by Western blot. Protein intracellular localization was shown by immunestaining and confocal microscopy. Results SH3P12 could precipitate and co-localize with FasL in 293T double transfectants. The interaction of SH3P12/FasL results in an enhanced surface expression of FasL. Conclusion SH3P12 is a FasL interacting protein; it might be involved in FasL surface expression.

关 键 词：FAS配体 FasL相关蛋白 蛋白质转运 蛋白-蛋白相互作用 

分 类 号：R392[医药卫生—免疫学]