Molecular mechanisms of Gram-negative outer membrane organization

革兰氏阴性外膜组织的分子机制

• 批准号: 10294945
• 负责人: Irina V. Mikheyeva-Bridges
• 金额: $ 5.37万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10294945
• 关键词: Address; Affect; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Bacterial Physiology; Biological Assay; Cell Membrane Permeability; Cell membrane; Cells; Cellular biology; Centers for Disease Control and Prevention (U.S.); Cessation of life; Communication; Complex; Crowding; Data; Development; Divalent Cations; Drug resistance; Educational process of instructing; Environment; Escherichia coli; Exclusion; Face; Fellowship; Fluorescence Microscopy; Frequencies; Genetic Techniques; Goals; Gram-Negative Bacteria; Health; Human; Hydrophobicity; Image Analysis; Infection; Island; Knowledge; Label; Lead; Lipid Bilayers; Lipopolysaccharides; Maintenance; Membrane; Membrane Proteins; Mentors; Modeling; Molecular; Molecular Biology; Monitor; Mutation; Nutrient; O Antigens; Organelles; Permeability; Phenotype; Philosophy; Phospholipids; Physiologic pulse; Property; Protein Dynamics; Proteins; Public Health; Research; Role; Running; Stress; Structure; Techniques; Total Internal Reflection Fluorescent; Training; United States; Universities; VDAC1 gene; Visualization; Work; antibiotic design; bacterial genetics; beta barrel; career; collaborative environment; design; experimental study; fungus; graduate student; in vivo; meetings; member; membrane model; microscopic imaging; mutant; novel; prevent; protein function; quantitative imaging; scaffold; skills; small molecule; symposium; tool; undergraduate student; uptake; wasting

The rising frequency of drug-resistant bacterial infections pose an imminent threat to human health. Attempts to generate novel antibiotics to address this problem often fail because the outer membrane (OM) of gram-negative bacteria acts as a barrier that blocks the entry of many small molecules. This OM barrier is densely packed with lipopolysaccharides (LPS) and outer membrane proteins (OMPs) that organize heterogeneously within the OM. Specifically, it has been demonstrated that OMPs form higher-order assemblies in the membrane termed “islands”. The OM porins are trimeric and highly abundant OMPs but it is unknown how this oligomerization state influences their function, insertion into the OM, or the assembly of OMP islands. I hypothesize that OMP trimerization is critical for OMP function and fundamentally underlies the heterogenous organization of the OM. Using the model OMP, LamB, I have developed a set of tools that will allow me to examine this hypothesis. First, I will use bacterial genetic techniques to determine how OMP oligomeric state influences OMP function and partitioning of LPS. Second, I will use fluorescence microscopy techniques to characterize the role of OMP oligomerization in the OMP island assembly. Finally, I will determine how local secretion of OMPs affect OM organization. Together, my proposed work will define the molecular mechanisms that govern the organization of the OM. Importantly, this will inform strategies for disrupting the OM and in designing OM permeable antibiotics. The training goal of this fellowship is to continue to grow my scientific knowledge while further developing skills as a leader and mentor in order to fulfill my career goal of becoming a research group leader. Being a member of the Silhavy lab at Princeton University will allow me to develop a teaching philosophy and participate in the teaching and mentoring of undergraduate and graduate students. I will perfect my communication skills through ample opportunities to present my work at meetings and conferences. Additionally, the Molecular Biology Department at Princeton University is collaborative, interdisciplinary environment that promotes that development of ideas and sharing of expertise. Together, I believe that the completion of this training plan in the research environment of the Silhavy lab at Princeton University will allow me to develop the necessary skills to run my own research group.

耐药细菌感染日益频繁，对人类健康构成迫在眉睫的威胁。