Structure and Assembly of Type IV Pili & Related Systems

IV型霹雳的结构与组装

• 批准号: 7214771
• 负责人: John A. Tainer
• 金额: $ 34.82万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214771
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Address; Adhesions; Amino Acid Sequence Homology; Antibiotic Resistance; Archaeoglobus fulgidus; Architecture; Binding; Biochemical; Biochemical Genetics; Biological; Biological Assay; Biological Process; Chemistry; Classification; Complex; Computer Simulation; Core Assembly; Crystallography; Cytoplasmic Tail; DNA; Data; Deuterium; Dichelobacter nodosus; Disease; Electron Microscopy; Enzymes; Epitopes; Fiber; Figs - dietary; Filament; Fimbriae Proteins; Flagella; Flagellin; Gram-Negative Bacteria; Helix (Snails); Higher Order Chromatin Structure; Homologous Gene; Imagery; Immune response; Immunoglobulin Variable Region; Individual; Investigation; Joints; Knowledge; Length; Mass Spectrum Analysis; Membrane; Membrane Proteins; Microbe; Microbial Biofilms; Modeling; Molecular; Molecular Conformation; Mutagenesis; N-terminal; Outcome; Pathogenesis; Pathogenicity; Pilum; Process; Proteins; Pseudomonas aeruginosa; Reagent; Research; Resolution; Role; Secretin; Signal Transduction; Site; Structure; Surface; System; Techniques; Terrorism; Tertiary Protein Structure; Testing; Therapeutic; Therapeutic Intervention; Toxin; Type II Secretion System Pathway; Vaccines; Variant; Vibrio cholerae; Virulence Factors; Work; X-Ray Crystallography; base; cell motility; design; drug discovery; ear helix; membrane assembly; novel vaccines; pathogen; periplasm; pill; protein protein interaction; protein structure; reconstruction; research study; scaffold; structural biology; therapeutic vaccine; three dimensional structure

DESCRIPTION (provided by applicant): A molecular understanding of bacterial virulence factors is critical to address the urgent need for effective new vaccines and therapeutics for increasingly dangerous antibiotic resistant microbes, emerging diseases and bio-terrorism threats. Type IV pill (T4P) are key virulence factors for Gram negative bacteria with diverse roles in surface motility, adhesion, microcolony and biofilm formation, signal transduction, and DNA transformation. These surface-exposed T4P are targets for the host immune response, as well as for vaccines and therapeutic reagents. The T4P system is structurally related to the Type II secretion system (T2SS), which exports toxins in pathogenic bacterial species, and to the archaeal flagellar system, needed for motility. These three systems employ multiple protein components to build multifunctional filaments spanning the periplasmic space. To achieve detailed characterizations of such challenging filament systems, we will integrate biochemical, biophysical and structural characterizations by both x-ray crystallography and electron microscopy. These experiments aim to solve a prototypical set of subunit and filament structures, and to define assembly ATPase conformations and their complexes with membrane protein partners. We will work outward from the subunit cores to the conformational states and interaction domains and then to characterize interfaces and protein:protein complexes. This joint Tainer-Craig proposal will thus provide an integrated visualization built up from the individual protein domains to the complex T4P and T2SS systems and thereby fill a critical gap in the structural biology of prokaryotic filamentous assemblies. Assembly structures will be further tested by mutagenesis and deuterium exchange-mass spectrometry. The proposed experiments will thus build a molecular description of assemblies relevant to understanding their pathogenic variation to escape the immune response, diverse biological roles, target sites for drug discovery, and design with protective epitopes for vaccines.

描述（由申请人提供）：对细菌毒力因子的分子理解对于解决日益危险的抗生素耐药微生物、新出现的疾病和生物恐怖主义威胁的有效新疫苗和疗法的迫切需求至关重要。IV型片剂（T4P）是革兰氏阴性菌的关键毒力因子，在表面运动、粘附、微菌落和生物膜形成、信号转导和DNA转化等方面发挥着不同的作用。这些表面暴露的T4P是宿主免疫反应的靶标，也是疫苗和治疗试剂的靶标。T4P系统在结构上与II型分泌系统（T2SS）和古细菌鞭毛系统有关，后者在致病菌中输出毒素，并与运动所需的古细菌鞭毛系统有关。这三个系统使用多种蛋白质成分来构建跨越质周空间的多功能细丝。为了实现这种具有挑战性的细丝系统的详细表征，我们将通过x射线晶体学和电子显微镜整合生化，生物物理和结构表征。这些实验旨在解决一组典型的亚基和丝结构，并定义组装atp酶构象及其与膜蛋白伙伴的复合物。我们将从亚基核心向外研究构象状态和相互作用域，然后表征界面和蛋白质：蛋白质复合物。因此，这个联合Tainer-Craig的提议将提供一个从单个蛋白质结构域到复杂的T4P和T2SS系统的集成可视化，从而填补了原核生物丝状组装结构生物学的关键空白。组装结构将通过诱变和氘交换质谱进一步测试。因此，拟议的实验将建立与了解其致病变异以逃避免疫反应相关的组装的分子描述，多种生物作用，药物发现的靶点，以及疫苗保护表位的设计。