Structural Mechanisms of MALT1 Signaling in T-Cell Differentiation and Activation (B02)

T 细胞分化和激活中 MALT1 信号传导的结构机制 (B02)

• 批准号: 230012849
• 金额: --
• 依托单位: Genzentrum - Laboratorium für molekulare Biologie
• 依托单位国家: 德国
• 项目类别: Collaborative Research Centres
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/230012849?language=en
• 关键词: Structural; Mechanisms; MALT1; Signaling; Cell

In the last funding period, we solved the architecture of the helical BCL10-MALT1 core filament by cryo-electron microscopy (cryo-EM), defining the interface between the N-terminal MALT1 death domain (DD) and the BCL10 caspase recruitment domain (CARD) and by showing that the C-terminal MALT1 immunoglobulin and paracaspase domains protrude flexibly from this inner core to orchestrate binding of mediators and substrates at the filament periphery. Based on these discoveries, we will now analyze the recruitment of TRAF6 to the C-terminus of MALT1 as well as substrate binding to MALT1 by biochemical techniques, X-ray crystallography and cryo electron microscopy. To account for its regulation by phosphorylation, mono- and poly-ubiquitination we will purify the CBM proteins from human cells with the goal to analyze posttranslational-modified samples by cryo-electron microscopy (cryo-EM) or individual filaments by cryo-electron tomography (cryo-ET). Finally, we will develop a method to investigate the CBM complex filament assembly in T cells using in cellulo cryo-ET.

在上一个资助期间，我们通过冷冻电子显微镜（cryo-EM）解决了螺旋BCL 10-MALT 1核心细丝的结构，定义了N-末端MALT 1死亡结构域（DD）和BCL 10半胱天冬酶募集结构域（CARD）之间的界面，并通过显示C-末端MALT 1免疫球蛋白和半胱天冬酶结构域从该内核灵活地突出，以协调介体和底物在末端的结合。花丝周边基于这些发现，我们现在将通过生物化学技术、X射线晶体学和冷冻电子显微镜分析TRAF 6向MALT 1的C末端的募集以及与MALT 1的底物结合。为了解释其通过磷酸化、单泛素化和多泛素化的调节，我们将从人类细胞中纯化CBM蛋白，目的是通过冷冻电子显微镜（cryo-EM）或通过冷冻电子断层扫描（cryo-ET）分析术后修饰的样品。最后，我们将开发一种方法来研究CBM复合物细丝组装在T细胞中使用在cellulocryo-ET。