Membrane Protein Structures and Interactions in the M. tuberculosis Divisome

结核分枝杆菌分裂体中的膜蛋白结构和相互作用

• 批准号: 8944802
• 负责人: TIMOTHY A CROSS
• 金额: $ 76.44万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-20 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8944802
• 关键词: Antibiotics; Bacillus subtilis; Bacteria; Binding; Binding Sites; Biology; Biophysics; C-terminal; Cell Wall; Cell division; Cell membrane; Cells; Cessation of life; Communication; Complex; Cytoplasm; Disease; Drug Targeting; Drug resistance in tuberculosis; Environment; Escherichia coli; Exposure to; Foundations; Goals; Homologous Gene; Individual; Integral Membrane Protein; Joints; Knowledge; Label; Lead; Length; Lipid Bilayers; Lipids; Magic; Membrane; Membrane Proteins; Methodology; Microbiology; Multiple Partners; Mycobacterium tuberculosis; Peptidoglycan; Pharmaceutical Preparations; Positioning Attribute; Process; Proteins; Publications; Recruitment Activity; Research; Sampling; Solutions; Staging; Structural Protein; Structure; Surface; Technology; Therapeutic; Toxin; Transmembrane Domain; Tuberculosis; Two-Hybrid System Techniques; Water; Work; daughter cell; experience; falls; frontier; interest; killings; meetings; molecular dynamics; novel; novel therapeutics; prevent; protein complex; protein protein interaction; protein structure; public health relevance; research study; restraint; solid solution; solid state; solid state nuclear magnetic resonance; structural biology; tool; tuberculosis drugs; tuberculosis treatment

 DESCRIPTION (provided by applicant): Relatively little is known about the cell division machinery, or divisome, of Mycobacteria tuberculosis (Mtb), the causative agent of tuberculosis (TB). There have been no new TB drug classes developed since a multidrug 6-month treatment course was introduced in the 1970s. New therapies are desperately needed for TB as the occurrence of multi-drug and extensive drug-resistant TB has increased rapidly in the past decade. The goal of this project is to advance knowledge of the Mtb divisome as potential drug targets and towards this end achieve an understanding of how the divisome proteins interact with each other and hence, how they are recruited to the divisome. Four transmembrane proteins are targeted for structural characterizations, both individually and as complexes. The Specific Aims are: (1) characterize nascent and binding-induced structures of intrinsically disordered regions in the protein targets; (2) determine full-length structures of these proteins i lipid bilayers; (3) determine structures of complexes formed between these membrane proteins and with other divisome proteins and peptidoglycan precursors or fragments. Combining expertise in microbiology and structural biology with membrane protein and computational biophysics, the research will lead to critical knowledge on the activities and interactions of central layers in the divisome. An array of tools will be used, including bacterial two-hybrid assays, solid state and solution NMR, and restrained molecular dynamics. To meet the challenge of characterizing the proteins and protein complexes in a native-like, lipid bilayer environment, new structural methodologies, including a novel type of structural restraints from solid state NMR and a novel use of solid state NMR for conformational characterization of intrinsically disordered regions in membrane proteins, will be developed. This interdisciplinary team, with significant and unique experiences of the individuals and through prior joint publications, is ideally positioned to accomplish the proposed studies. These structural characterizations of the Mtb divisome and their functional implications will generate novel therapeutic opportunities for TB. The technologies developed here will represent the frontier of membrane protein structure biology. 1

 描述（由申请人提供）：关于结核病（TB）的病原体结核分枝杆菌（Mtb）的细胞分裂机制或分裂体的了解相对较少。自20世纪70年代采用6个月多药疗程以来，没有开发出新的结核病药物类别。在过去十年中，由于多药耐药和广泛耐药结核病的发生率迅速增加，迫切需要新的结核病疗法。该项目的目标是推进Mtb divisome作为潜在药物靶点的知识，并为此实现对divisome蛋白如何相互作用以及它们如何被招募到divisome的理解。四个跨膜蛋白的结构表征，无论是单独的和作为复合物的目标。具体目标是：（1）表征蛋白质靶中固有无序区域的新生和结合诱导的结构;（2）确定脂质双层中这些蛋白质的全长结构;（3）确定这些膜蛋白之间以及与其它分裂体蛋白和肽聚糖前体或片段形成的复合物的结构。将微生物学和结构生物学的专业知识与膜蛋白和计算生物物理学相结合，这项研究将导致对分裂中中心层的活动和相互作用的关键知识。将使用一系列工具，包括细菌双杂交测定，固态和溶液NMR，和限制分子动力学。为了满足的挑战，表征蛋白质和蛋白质复合物在一个天然的，脂质双层环境，新的结构方法，包括一种新的类型的结构限制从固态NMR和一种新的使用固态NMR的固有的膜蛋白质中的无序区域的构象表征，将开发。这个跨学科的团队，具有重要和独特的个人经验，并通过以前的联合出版物，是理想的定位，以完成拟议的研究。结核分枝杆菌分裂体的这些结构特征及其功能意义将产生新的治疗结核病的机会。这里开发的技术将代表膜蛋白结构生物学的前沿。1