Composition stoichiometry of CD95/Fas complexes as a switch between life and death

细胞程序性死亡是一个高度调控的过程，对所有多细胞生物体都是必不可少的。放松对细胞死亡的管制会导致许多疾病，如癌症、神经退行性疾病和自身免疫性疾病。有两种主要类型的细胞程序性死亡：细胞凋亡和坏死下垂，它们可以通过连接死亡受体(DR)而启动。CD95/Fas/APO-1是DR家族中研究最深入的成员之一。据报道，CD95的刺激不仅导致细胞死亡，而且还诱导了一系列的生存/增殖途径，如核因子-kB。一个特定的CD95通路的诱导依赖于相应信号平台或多蛋白复合体的形成。该提案将致力于分析化学计量学、翻译后修饰(PTM)和寻找CD95分子平台的新分子，调节细胞凋亡、坏死性下垂和核因子-kB之间的开关。我们将在我们的工作中使用高度创新的方法：从当代的质谱分析到单细胞测量。络合物的分离将遵循该领域中确立的技术状态，并将涉及高分子量分馏。我们的研究将为CD95网络调控的分子机制提供新的信息，并阐明这些信号平台在疾病背景下如何潜在地成为靶点的问题。

Programmed cell death is a highly regulated process which is essential for all multicellular organisms. The deregulation of cell death leads to a number of diseases such as cancer, neurodegenerative and autoimmune diseases. There are two major types of programmed cell death: apoptosis and necroptosis that could be initiated upon ligation of death receptors (DR). CD95/Fas/APO-1 is one of the best-studied members of the DR family. Stimulation of CD95 has been reported to result not only in the cell death but also in the induction of a number of survival/proliferation pathways such as NF-kB. The induction of a particular CD95 pathway is crucially dependent on the formation of the corresponding signaling platform or multiprotein complex. The proposal will be devoted to the analysis composition stoichiometry, posttranslational modifications (PTMs) and search for new molecules of CD95 molecular platforms regulating the switch between apoptosis, necroptosis and NF-kB. We will use highly innovative approaches in our work: from the contemporary Mass Spectrometry analysis to the single cell measurements. The isolation of the complexes will be followed to the established state of the art in the field and would involve high molecular weight fractionation. Our studies should provide a new information on molecular mechanisms of the CD95 network regulation and shed the light on the question how these signaling platforms could potentially be targeted in the context of disease.