Mechanism of membrane fusion involving the Gram-negative bacteria outer membrane

涉及革兰氏阴性菌外膜的膜融合机制

基本信息

批准号：
10089745
负责人：
Reza Khayat
金额：
$ 37.83万
依托单位：
CITY COLLEGE OF NEW YORK
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10089745
关键词：
Amino Acid Sequence Amino Acid Substitution Antibiotic Resistance Antibiotic-resistant organism Antibiotics Architecture Area Bacteriophages Binding Biochemical Bioinformatics Biophysics Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Communication Complex Cryo-electron tomography Cryoelectron Microscopy Cystovirus Data Escherichia coli Eukaryota Eukaryotic Cell Exclusion Family Family member Fluorescence Spectroscopy Glycoproteins Goals Gram-Negative Bacteria HIV Health Herpesviridae Horizontal Gene Transfer Human Infection Knowledge Lateral Life Light Lipid A Lipopolysaccharides Membrane Membrane Fusion Modernization Molecular Conformation Molecular Machines Multiprotein Complexes Muscle Cells Myoblasts Nature Nerve O Antigens Organelles Pathogenicity Peptides Permeability Phospholipids Process Property Proteins Pseudomonas aeruginosa Pseudomonas syringae Public Health Reporting Research Salmonella typhimurium Site Structure Synaptic Vesicles System Technology Testing Vacuole Vesicle Viral Virus Diseases West Nile virus X-Ray Crystallography antibiotic resistant infections bacterial resistance biophysical analysis egg member nanotherapeutic pathogen pathogenic bacteria pathogenic fungus protein complex recruit scaffold sperm cell

项目摘要

PROJECT SUMMARY/ABSTRACT Gram-negative bacteria constitute the majority of antibiotic resistant organisms identified as urgent threats to human health by the Center for Disease Control and Prevention. A major reason why Gram-negative bacteria are resistant to modern antibiotics is the impermeable nature of their outer membrane lipopolysaccharide (LPS) layer. This impermeable nature is due to the strong lateral interaction between neighboring LPS molecules that includes lipid A, the core saccharide, and the O-antigen. Nanotherapeutics capable of overcoming this barrier to successfully deliver antibiotics to Gram-negative bacteria will be of immense importance for human health. It is our intent to develop such nanotherapeutics by using the knowledge attained from delineating the mechanism by which a Gram-negative bacteriophage fuses its external bacterial phospholipid (BPL) membrane with the LPS of its host for initiating infection. Embedded into the Cystovirus BPL are multimeric protein complexes, referred to as spikes, that recognize the Gram-negative host and perform fusion. These proteins are analogous to, but different from, the better studied eukaryotic proteins responsible for membrane fusion (e.g. HIV env, West Nile Virus glycoprotein E, and Herpesviridae gB protein). The intent of this proposal is to investigate the mechanism by which spikes from three different members of the Cystovirus family recognize their hosts and drive membrane fusion. The goals of this proposal are to determine the structure of the spikes for establishing a structural scaffold for biochemical and biophysical studies (Aim 1), determine the mechanism of cellular recognition (Aim 2), and determine the biochemical determinants responsible for LPS- BPL fusion (Aim 3). The long-term goal of this project is to use the spikes for delivery of antibiotics to specific strains of pathogenic Gram- negative bacteria.

项目概要/摘要革兰氏阴性菌占抗生素抗性微生物的大多数，被认为是对抗生素的紧急威胁人类健康由疾病控制和预防中心负责。革兰氏阴性菌产生的主要原因对现代抗生素具有耐药性的原因在于其外膜脂多糖 (LPS) 的不渗透性层。这种不可渗透的性质是由于相邻 LPS 分子之间强烈的横向相互作用造成的包括脂质A、核心糖和O-抗原。纳米疗法能够克服这一障碍成功地将抗生素传递给革兰氏阴性菌对于人类健康至关重要。这是我们打算利用从描述机制中获得的知识来开发这种纳米疗法革兰氏阴性噬菌体将其外部细菌磷脂 (BPL) 膜与其宿主的LPS用于引发感染。嵌入囊病毒 BPL 中的是多聚蛋白复合物，称为刺突，可识别革兰氏阴性宿主并进行融合。这些蛋白质是类似的与更好地研究的负责膜融合的真核蛋白（例如 HIV env、West 尼罗病毒糖蛋白 E 和疱疹病毒 gB 蛋白）。该提案的目的是调查来自囊病毒家族三个不同成员的尖峰识别其宿主的机制驱动膜融合。该提案的目标是确定尖峰的结构，以建立用于生化和生物物理研究的结构支架（目标 1），确定细胞的机制识别（目标 2），并确定负责 LPS-BPL 融合的生化决定因素（目标 3）。这该项目的长期目标是使用尖峰将抗生素输送到特定的致病性革兰氏菌菌株阴性菌。