Protein Interactions in the Flagellar Export Machinery

鞭毛输出机械中的蛋白质相互作用

• 批准号: 6893385
• 负责人: JONATHAN L MCMURRY
• 金额: $ 4.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6893385
• 关键词: Salmonella typhimurium; analytical ultracentrifugation; bacterial proteins; cilium /flagellum motility; flagellum; fluorescence resonance energy transfer; fluorescence spectrometry; green fluorescent proteins; ion exchange chromatography; membrane biogenesis; membrane proteins; microorganism growth; molecular assembly /self assembly; postdoctoral investigator; protein protein interaction; protein purification; protein structure function; stoichiometry; western blottings

DESCRIPTION (provided by applicant): Motility plays a critical role in bacterial pathogenesis. Many species utilize flagella propulsion to locate hosts and move through tissues. The flagellum is a complex, reversible, self-assembling nanomachine. It contains its own type III protein export machinery that is closely related to the virulence factor secretion apparatus. The membrane proteins of the export apparatus are probably the least understood components of the flagellum. The overall objective of the proposed research is to obtain detailed knowledge of the interactions among these proteins from Salmonella typhimurium. Additionally, one of the ultimate goals of efforts to understand flagellar assembly is to reassemble the flagellar export complex from components to determine and analyze the roles of constituent proteins as well as the machinery itself. In order to achieve the specific objectives of the proposal and to approach the long term goal, several efforts will be undertaken: identification of interactions among the membrane proteins within the export apparatus (FIhA, FlhB, FliO, FliP, FliQ and FliR) and between the membrane components and other flagellar proteins by biochemical methods such as affinity blotting, copurification, etc.; biophysical analysis of interactions with analytical ultracentrifugation and fluorescence resonance energy transfer studies; and reconstitution of complexes of export apparatus proteins and analysis of their concerted function.

描述（由申请方提供）：运动性在细菌发病机制中起关键作用。许多物种利用鞭毛推进来定位宿主并在组织中移动。鞭毛是一种复杂的、可逆的、自组装的纳米机器。它含有自己的III型蛋白质输出机制，与毒力因子分泌装置密切相关。输出器的膜蛋白可能是鞭毛中最不为人所知的成分。拟议研究的总体目标是获得这些蛋白质之间的相互作用的详细知识，从鼠伤寒沙门氏菌。此外，努力了解鞭毛组装的最终目标之一是从组件中重新组装鞭毛输出复合物，以确定和分析组成蛋白质以及机器本身的作用。为了实现该提案的具体目标并接近长期目标，将进行几项努力：通过生物化学方法如亲和印迹、共纯化等鉴定输出装置内的膜蛋白（FIhA、FlhB、FliO、FliP、FliQ和FliR）之间以及膜组分与其他鞭毛蛋白之间的相互作用;与分析性超离心和荧光共振能量转移研究的相互作用的生物物理分析;以及输出装置蛋白质的复合物的重建和它们的协同功能的分析。