Secretin Architecture

分泌素架构

• 批准号: 9245656
• 负责人: MICHAEL S DONNENBERG
• 金额: $ 19.19万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9245656
• 关键词: Architecture; Bacteria; Biochemical; Biogenesis; Biological Models; Biomedical Research; Cells; Complex; Cryoelectron Microscopy; Cysteine; Development; Disease; Disulfides; Electron Microscopy; Energy Transfer; Equipment; Exploratory/Developmental Grant; Fiber; Fimbriae Proteins; Goals; Homologous Gene; Human; Imagery; Ion Channel; Knowledge; Length; Maps; Membrane; Microscopic; Modeling; Mutagenesis; Pilum; Positioning Attribute; Proteins; Publishing; Research; Research Project Grants; Resolution; Secretin; Shapes; Site; Structure; Surface; System; Techniques; Technology; Toxin; Virulence Factors; X-Ray Crystallography; crosslink; disulfide bond; electron tomography; enteropathogenic Escherichia coli; experimental study; fluorophore; improved; in vivo; monomer; mutant; novel; novel therapeutics; periplasm; permissiveness; protein complex; public health relevance; tool

 DESCRIPTION (provided by applicant): The secretin, a homomultimeric outer membrane channel, is the only protein common to type IV pilus (T4P), type 2 secretion and type 3 secretion systems. Therefore, secretins enjoy a unique position of prominence among virulence factors. Although the structures of several periplasmic secretin domains have been solved by X-ray crystallography, and the overall shape of secretins has been described by electron microscopy, sufficient detail to understand the mechanism that allows secretins to interact with other components of their respective machines and to open selectively to allow passage of their substrates has not be achieved. In this proposal we describe how we will advance our understanding of secretin function. Using a variety of techniques, including cysteine mutagenesis and Förster resonance energy transfer, we described a topology model for BfpB, the secretin from the T4P of enteropathogenic Escherichia coli. The first aim of this proposal will build on that knowledge by using in vivo disulfide cross-linking to trap the pilin in the secretin and thereby map the walls of the secretin channel. Recent advances in cryo-electron microscopy have permitted the structural analysis of large complexes at resolutions approaching the atomic range. In aim two we will capitalize on these advances to visualize the secretin and proteins with which it associates in unprecedented detail. The experiments described in this proposal will provide critical structural information that is essential for our lng-term goal of a complete understanding of T4P biogenesis. Furthermore, this information will also have implications for secretion systems and may ultimately have practical implications for new therapeutics.

 描述（由申请人证明）：Systrin，一种同性恋外膜通道，是唯一的蛋白质通用打字（T4P）和键入3个分泌系统，因此，Secitions在毒力中具有突出性的独特位置。通过电子显微镜描述的X射线晶体学，Severrasic Sextin域已求解，这是允许Cretins与其Tive机的组件相互作用的机制，并有选择地允许其底物通过。 在第二个二硫化物的二硫键交联的基础上，他们通过绘制了sectionin渠道的绘制。利用这些进展，可将其与前所未有的细节相关联的分泌素和蛋白质。同样对安全的影响，最终对治疗学具有实际影响。