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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 study the effect of FasL-related protein SH3P12 on FasL expression. Methods SH3P12 eukaryotic expression vector was constructed and transiently expressed in human embryonic kidney 293T cells with FasL. The co-immunoprecipitation and immunoblotting were used to determine the interaction between the target proteins. The expression of SH3P12 and FasL was observed by immunofluorescence staining and confocal laser scanning microscopy. Results In transfected 293T cells, SH3P12 and FasL formed an immunoprecipitated protein-protein complex. Fluorescent protein labeling combined with microscopy suggested that in the absence of SH3P12, FasL was mainly distributed in the 293T cell membrane with some proteins expressed in intracellular Golgi or inclusion body-like particles; in the presence of SH3P12, FasL formed a complex with SH3P12 , Co-located in the cell membrane, when the amount of intracellular expression of FasL was significantly reduced. Conclusion SH3P12 is a FasL intracellular binding protein and its biological activity may be the expression of stable FasL on the cell membrane.