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

• 批准号: 10400637
• 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
• 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/10400637
• 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

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.

项目摘要/摘要 革兰氏阴性细菌构成大多数抗生素耐药的生物，被确定为紧急威胁 疾病控制与预防中心的人类健康。革兰氏阴性细菌的主要原因 对现代抗生素具有抵抗力是其外膜脂多糖（LPS）的不渗透性质 层。这种不可渗透的性质是由于相邻LPS分子之间的强烈横向相互作用引起的 包括脂质A，核糖和O-抗原。纳米疗法能够克服这个障碍 成功地将抗生素传递给革兰氏阴性细菌对人类健康至关重要。这是 我们使用划定机制所获得的知识来开发这种纳米疗法的意图 革兰氏阴性噬菌体将其外部细菌磷脂（BPL）膜融合 其宿主的LPS发起感染。嵌入半胱氨酸病毒BPL的是多聚体蛋白质复合物， 称为尖峰，可以识别革兰氏阴性宿主并进行融合。这些蛋白质类似 to，但与不同的研究的真核蛋白负责膜融合（例如HIV Env，West， 尼罗河病毒糖蛋白E和疱疹病毒GB蛋白）。该提议的目的是调查 圣单鼠家庭三个不同成员的尖峰的机制认识了他们的宿主和 驱动膜融合。该提案的目标是确定建立尖峰的结构 生化和生物物理研究的结构支架（AIM 1）确定细胞的机制 识别（AIM 2），并确定负责LPS-BPL融合的生化决定因素（AIM 3）。这 该项目的长期目标是利用尖峰将抗生素递送到特定的致病性革命菌株中 阴性细菌。