Scaffidi C, Krammer P H, Peter M E
Tumor Immunology Program, German Cancer Research Center, Im Neuenheimer Feld 280, Heidelberg, 69120, Germany.
Methods. 1999 Apr;17(4):287-91. doi: 10.1006/meth.1999.0742.
CD95 (APO-1/Fas) is an apoptosis-inducing receptor belonging to the tumor necrosis factor receptor superfamily. Multimerization of CD95 leads to instant recruitment of the signaling molecules FADD and caspase-8 to the activated receptor forming the death-inducing signaling complex (DISC). DISC formation is the first essential step of CD95 signaling and results in activation of caspase-8 starting a signaling cascade that leads to apoptosis. Here we describe a method for analyzing the CD95 DISC. The method is based on coimmunoprecipitation of the signaling molecules with the activated CD95 receptor followed by Western blot detection of associated molecules. Therefore, this method can analyze the very first signaling events during CD95-mediated apoptosis.
CD95（APO-1/Fas）是一种属于肿瘤坏死因子受体超家族的凋亡诱导受体。CD95的多聚化导致信号分子FADD和半胱天冬酶-8立即募集到活化的受体上，形成死亡诱导信号复合物（DISC）。DISC的形成是CD95信号传导的第一个关键步骤，导致半胱天冬酶-8激活，启动导致细胞凋亡的信号级联反应。在此，我们描述一种分析CD95 DISC的方法。该方法基于信号分子与活化的CD95受体的免疫共沉淀，随后通过蛋白质印迹法检测相关分子。因此，该方法可以分析CD95介导的细胞凋亡过程中最初的信号事件。
