Assembly and Function of the Bacterial Type III Secretion System Basal Body

细菌III型分泌系统基体的组装和功能

• 批准号: 8535920
• 负责人: Charles Erec Stebbins
• 金额: $ 42.38万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535920
• 关键词: Animals; Anti-Bacterial Agents; Bacteria; Bacterial Proteins; Bacterial Toxins; Bacterial Typing; Biology; Cells; Chemicals; Collaborations; Communicable Diseases; Complex; Development; Devices; Disease; Electron Microscopy; Engineering; Ensure; Escherichia coli; Eukaryotic Cell; Gram-Negative Bacteria; Grant; Hand; Health; Human; Image; Indium; Infection; Injection of therapeutic agent; Integral Membrane Protein; Investments; Laboratories; Learning; Medicine; Membrane; Methods; Molecular; Molecular Models; Nanotechnology; Organism; Physiology; Plants; Process; Production; Proteins; Protocols documentation; Quality Control; Recombinants; Research; Research Personnel; Resolution; Risk; Roentgen Rays; Salmonella; Science; Screening procedure; Shigella; Solutions; Structure; Syringes; System; Techniques; Therapeutic; Time; Type III Secretion System Pathway; Universities; Virulence; Virulence Factors; Work; X-Ray Crystallography; base; design; fascinate; genetic manipulation; kinetosome; microorganism; milligram; molecular modeling; nanodevice; nanomachine; novel; pathogen; periplasm; programs; protein structure function; success

DESCRIPTION (provided by applicant): The protein Type III Secretion System (T3SS) is a supramolecular, organic nanomachine that injects bacterial virulence proteins into eukaryotic cells to modulate their physiology for the benefit of the pathogen. It underlies the virulence of numerous medically relevant organisms. A mechanistic understanding of this system is one of the central scientific problems in modern infection biology. At the same time, it is a problem whose solution will also provide important translational benefits, allowing for the development of antivirulence compounds, which target the injection device. Furthermore, the T3SS represents one of the most fascinating examples of a nanomachine. Intellectually, studying the structure, function, and mechanism of the T3SS will impact the understanding of infectious diseases in animals and plants, as well as increase the understanding of nanotechnology and machineries. This grant proposes to solve a specific problem: what is the structural basis for the assembly and function of the T3SS spanning the two membranes of Gram negative bacteria? The project is divided into three specific aims: (1) Optimize and extend current protocols for the expression and purification of T3SS syringe complexes (2) Develop the first recombinant methods for expressing and purifying the secretion complex, and (3) Determine the high-resolution structure of the T3SS nanomachine by X-ray crystallography. To understand the T3SS at the molecular level, high-resolution structural information is required. While moderate resolution electron microscopy (EM) has given an overall view of the system, X-ray crystallographic studies will be required to develop a mechanistic understanding. Such a "blueprint" for the assembly of the T3SS will be a critical first step towards understanding its function and how to inhibit this function in a therapeutic context.

描述(申请人提供)：蛋白III型分泌系统(T3SS)是一种超分子有机纳米机器，它将细菌毒力蛋白注入真核细胞，以调节它们的生理，从而有利于病原体。它是许多与医学相关的生物体毒力的基础。从机制上理解这一系统是现代感染生物学的中心科学问题之一。与此同时，这是一个问题，其解决方案也将提供重要的翻译好处，允许开发针对注射设备的抗毒力化合物。此外，T3SS代表了纳米机器最吸引人的例子之一。在智力上，研究T3SS的结构、功能和机制将影响对动植物传染病的理解，也将增加对纳米技术和机械的理解。这项拨款旨在解决一个具体的问题：跨越革兰氏阴性细菌两层膜的T3SS的组装和功能的结构基础是什么？该项目分为三个具体目标：(1)优化和扩展现有的T3SS注射器复合体的表达和纯化方案；(2)开发第一个表达和纯化分泌物复合体的重组方法；(3)通过X射线结晶学确定T3SS纳米机器的高分辨率结构。为了在分子水平上理解T3SS，需要高分辨率的结构信息。虽然中分辨率电子显微镜(EM)已经给出了该系统的总体情况，但需要进行X射线结晶学研究来从机理上进行理解。这种组装T3SS的“蓝图”将是理解其功能以及如何在治疗背景下抑制这一功能的关键的第一步。