Membrane protein effectors of pathogen interactions with host

病原体与宿主相互作用的膜蛋白效应子

基本信息

批准号：
9071833
负责人：
Francesca M Marassi
金额：
$ 75.21万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9071833
关键词：
Antibiotic Resistance Antibiotics Bacteria Bacterial Adhesion Biological Cell Adhesion Cell membrane Cells Classification Complement Complex Computing Methodologies Data Detergents Engineering Environment Goals Health Human Immune system Immunity Integral Membrane Protein Ligands Lipid Bilayers Mediating Medical Membrane Membrane Proteins Methods Molecular NMR Spectroscopy Pharmacologic Substance Phospholipids Plague Play Proteins Research Role Sampling Structure Vaccines Virulence Factors Work Yersinia pestis bacterial resistance biothreat design drug development extracellular human mortality pathogen programs public health relevance targeted treatment therapeutic development three dimensional structure

Antibiotic Resistance Antibiotics Bacteria Bacterial Adhesion Biological Cell Adhesion Cell membrane Cells Classification Complement Complex Computing Methodologies Data Detergents Engineering Environment Goals Health Human Immune system Immunity Integral Membrane Protein Ligands Lipid Bilayers Mediating Medical Membrane Membrane Proteins Methods Molecular NMR Spectroscopy Pharmacologic Substance Phospholipids Plague Play Proteins Research Role Sampling Structure Vaccines Virulence Factors Work Yersinia pestis bacterial resistance biothreat design drug development extracellular human mortality pathogen programs public health relevance targeted treatment therapeutic development three dimensional structure

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项目摘要

DESCRIPTION (provided by applicant): The overall goal of this project is to develop a framework for understanding the way in which membrane proteins expressed on pathogen cell membranes mediate the interactions of pathogens with their human host. We focus on Ail (attachment invasion locus), a protein expressed in the outer cell membrane of the bacterium Yersinia pestis, which is the causative agent of plague. The Y. pestis bacterium is highly pathogenic, spreads rapidly and causes extremely high human mortality. Although it is sensitive to a restricted panel of antibiotics, the potential weaponization of aerolized bacteria with bio-engineered antibiotic resistance and the lack of an effective vaccine or therapy are major concerns, contributing to its classification as a Tier 1 Biothreat Agent. Ail is a key bacterial virulence factor and a prime candidate for therapeutic development due to its functions in mediating bacterial adhesion to host cells, and promoting bacterial resistance to human immunity. This project focuses on elucidating the molecular mechanisms of these key functions of Ail by determining its three-dimensional structure and structurally characterizing the interactions of Ail with its human ligands. These studies will focus on samples of Ail and its complexes incorporated in detergent-free lipid bilayer membranes because the biological activity of Ail is dramatically altered in the presence of micellar detergents. Furthermore, these studies will utilize NMR spectroscopy, a method adept at characterizing both structure and dynamics, because the extracellular loops of Ail that mediate the interactions with host are highly dynamic. Finally, these studies will complement the experimental data generated for Ail in lipid bilayer membranes, by developing computational methods tailored specifically for NMR structural analysis of integral membrane proteins within their functional environment of the phospholipid bilayer membrane. The methods will be designed to facilitate structure determination and increase structural quality, and they will be integrated with widely used programs for programs for NMR structure calculations so as to broaden the impact of our work.

描述（由适用提供）：该项目的总体目标是开发一个框架，以了解膜蛋白在病原体上表达的方式介导病原体与其人类宿主的相互作用。我们专注于AIL（固定侵袭基因座），这是一种在鼠疫耶尔西尼亚细菌的外细胞膜中表达的蛋白质，该蛋白是鼠疫的致病药物。 Y. Pestis细菌具有高度致病性，迅速传播，并引起极高的人类死亡率。尽管它对受限的抗生素敏感，但具有生物工程抗生素耐药性和缺乏有效的疫苗或治疗的潜在武器化武器化是主要的关注点，这有助于将其分类为层次1生物疗法。 AIL是一种关键的细菌病毒因子，也是治疗性发育的主要候选者，因为它在介导细菌粘附对宿主细胞的功能以及促进了对人类免疫力的耐药性。该项目的重点是阐明AIL这些关键功能的分子机制，通过确定其三维结构，并在结构上表征AIL与其人体配体的相互作用。这些研究将重点放在AIL及其复合物的样品中，因为在胶束存在的存在下，在胶束的存在下，AIL的生物学活性发生了巨大变化，因此将AIL及其复合物的样品纳入确定。此外，这些研究将利用NMR光谱，这是一种擅长表征结构和动力学的方法，因为介导与宿主相互作用的AIL的细胞外回路具有高度动态性。最后，这些研究将完成脂质双层膜中AIL生成的实验数据，通过开发专门针对磷脂双层膜功能性环境内的积分膜蛋白的NMR结构分析量身定制的计算方法。这些方法将旨在促进结构确定并提高结构质量，并将它们与广泛使用的NMR结构计算程序的程序集成，从而扩大我们的工作影响。