Structure-function analysis of mammalian formins FMNL1 and FMNL2

哺乳动物formins FMNL1和FMNL2的结构-功能分析

• 批准号: 170435620
• 负责人: Professor Dr. Matthias Geyer
• 金额: --
• 依托单位: Institut für Strukturbiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2018-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/170435620?language=en
• 关键词: Structure; function; analysis; mammalian; formins

Regulation of the actin cytoskeleton is a key process for the stability and motility of eukaryotic cells. A diverse family of formin proteins and WH2 domain containing proteins have recently been recognized as actin nucleators and potent polymerization factors of actin filaments. Formins are defined by the presence of a formin homology 2 (FH2) domain, yet, the modular domain architecture appears significantly different for the eight formin families found in man. This research project focuses on the biochemical and structural analysis of the regulatory domains of mammalian formins. The formin-related gene in leukocytes 2 (FMNL2) is myristoylated, an N-terminal lipid modification which targets a protein to cellular membranes. In preliminary work, we have established an in vitro myristoylation assay that allows characterization of FMNL2 in its native form. We plan to analyse how activation by triphosphate Cdc42 influences the conformation of the myristate in the presence of liposomes. Structural analysis of the lipidated protein will be performed and the interaction with the C-terminal autoregulation domain analysed. Also, we are able to purify a complex of the N- and C-terminal domains of FMNL2 which constitute a double homodimer of 58 kDa and 54 kDa. Finally, functional experiments will address the impact of tropomyosin on filament stabilization during elongation.

肌动蛋白细胞骨架的调节是真核细胞稳定和运动的关键过程。近年来，一个新的蛋白质家族被认为是肌动蛋白成核因子和肌动蛋白丝聚合因子。形成蛋白是由一个FH 2结构域的存在下定义的，然而，模块结构域的架构似乎显着不同的八个FH 2家族在man. This研究项目的重点是哺乳动物形成蛋白的调节域的生化和结构分析。白细胞中的formin相关基因2（FMNL 2）是豆蔻酰化的，这是一种将蛋白质靶向细胞膜的N-末端脂质修饰。在初步工作中，我们已经建立了一个体外豆蔻酰化测定，允许FMNL 2在其天然形式的表征。我们计划分析三磷酸Cdc 42的激活如何影响脂质体存在下肉豆蔻酸酯的构象。将对脂化蛋白进行结构分析，并分析与C末端自动调节结构域的相互作用。此外，我们能够纯化FMNL 2的N-和C-末端结构域的复合物，其构成58 kDa和54 kDa的双同源二聚体。最后，功能实验将解决的影响，原肌球蛋白对长丝的稳定性在延长。