STRUCTURAL BASIS OF AMINORABINOSE BIOSYNTHESIS LINKED TO POLYMYXIN RESISTANCE

与多粘菌素抗性相关的氨基阿拉伯糖生物合成的结构基础

• 批准号: 10238086
• 负责人: Vasileios I Petrou
• 金额: $ 24.9万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-13 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238086
• 关键词: Active Sites; Address; Anabolism; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Award; Bacteria; Bacterial Genome; Binding; Biology; Biomedical Technology; Catalysis; Cations; Chemicals; Chimera organism; Collection; Communities; Cryoelectron Microscopy; Crystallization; Development Plans; Docking; Dolichol Phosphates; Educational process of instructing; Electron Microscopy; Electron Spin Resonance Spectroscopy; Electrostatics; Environment; Enzymes; Eukaryota; Faculty; Family; Future; Genes; Genomic approach; Goals; Gram-Negative Bacteria; Grant; Health; Host Defense; Human; Incidence; Infection; Knowledge; Laboratories; Lead; Ligands; Link; Lipid A; Lipid Binding; Lipids; Lipopolysaccharides; Location; Membrane; Membrane Lipids; Membrane Proteins; Mentors; Methodology; Modification; Molecular; Molecular Conformation; Monosaccharides; Multi-Drug Resistance; Mutagenesis; Names; Orthologous Gene; Pathway interactions; Peptides; Phase; Polymyxin Resistance; Polymyxins; Polysaccharides; Positioning Attribute; Principal Investigator; Probability; Professional Competence; Reaction; Research; Research Proposals; Resistance development; Resort; Resources; Salmonella; Site; Structure; Therapeutic; Training; Transferase; United States National Institutes of Health; Universities; World Health Organization; Writing; X-Ray Crystallography; analog; antimicrobial drug; career; career development; design; enzyme structure; experience; flexibility; global health; glycosyltransferase; inorganic phosphate; member; multidisciplinary; particle; periplasm; programs; responsible research conduct; screening; structural biology; structural genomics; sugar; technology development; tenure track

Project Summary/Abstract The research proposal outlined in this application is consisted of two components designed to facilitate the transition of the principal investigator (PI) to an independent research career, a career development plan and a research plan. The PI has a multidisciplinary background, and through this proposal, seeks to finalize his training in Structural Biology of Membrane Proteins. The career development plan comprises a structured educational experience for the first two years of the award (K99 phase) that includes training in Electron Microscopy and Electron Paramagnetic Resonance (EPR) Spectroscopy for the study of Membrane Proteins, and enhancement of career skills, such as grant writing, laboratory management, teaching and responsible conduct of research. It includes a clear and actionable plan for identifying and successfully competing for an independent tenure-track faculty position by the end of the K99 phase. The PI has assembled a multi- disciplinary team of mentors, advisors and collaborators that will oversee and guide his training, research program and transition to independence. The research plan spans both the mentored (K99) and independent (R00) phases of the award. It involves mechanistic studies of the aminoarabinose sugar biosynthetic pathway in Gram-negative bacteria that is linked to resistance to polymyxin-class antibiotics, our last line of defense against multi-drug resistant infections. The research program for the K99 phase aims to build on knowledge gained from a recent solution structure of an enzyme in the aminoarabinose pathway, named ArnT, by the PI and his colleagues. The independent (R00) research program then aims to extend the structural studies to other transmembrane enzymes of the aminoarabinose pathway, working towards a complete structural description and functional characterization of the pathway. The core questions that the research program aims to address are: (1) How does the enzyme ArnT accommodate its two lipidic substrates within the context of its fold? (2) What are the molecular determinants that impart stereospecificity to some members of the ArnT enzyme family but not others? (3) How is the ArnT enzyme changing during substrate binding and catalysis? (4) How do the other transmembrane enzymes in the pathway look and how do they accomplish their functions? (5) How do other transmembrane enzymes in the pathway accommodate the lipidic sugar carrier undecaprenyl phosphate? To address these questions, the PI has put together a comprehensive research plan that utilizes state-of-the-art methodologies. Moreover, the plan includes some technology development aimed at extending the capabilities of electron microscopy within the context of the research program, and which, upon completion of the program, may be applicable to other research programs. The proposed studies for the K99 phase will largely take place in the prominent environment of Columbia University, which harbors a vibrant structural biology community. This environment is certain to facilitate the PI in the successful completion of the proposed research program and achieving his goal of transitioning into an independent research career.

项目摘要/摘要 本应用程序中概述的研究建议包括两个组成部分，旨在促进 首席研究员（PI）过渡到独立研究职业，职业发展计划和 研究计划。 PI具有多学科背景，通过该提案，试图完成他的 膜蛋白结构生物学的培训。职业发展计划包括一个结构化的 奖项的前两年（K99阶段）的教育经验，其中包括电子培训 显微镜和电子顺磁共振（EPR）光谱，用于研究膜蛋白， 并提高职业技能，例如赠款写作，实验室管理，教学和负责任 进行研究。它包括一个明确，可行的计划，用于识别并成功争夺 在K99阶段结束时，独立的终身教师职位。 PI组装了一个多 导师，顾问和合作者的纪律团队，将监督和指导他的培训，研究 计划和过渡到独立性。研究计划涵盖了指导者（K99）和独立 （R00）奖项的阶段。它涉及氨基酸糖糖生物合成途径的机械研究 在革兰氏阴性细菌中，与多聚染色蛋白级抗生素有关，这是我们的最后一条防御 针对多药抗性感染。 K99阶段的研究计划旨在以知识为基础 从PI的氨基酸途径中的酶途径中从最新的酶结构中获得的溶液结构。 和他的同事。然后，独立（R00）研究计划旨在将结构研究扩展到 氨基氨基酸途径的其他跨膜酶，致力于完整的结构 路径的描述和功能表征。研究计划旨在的核心问题 要解决 折叠？ （2）赋予某些ARNT成员的立体特定性的分子决定因素是什么 酶家族，但不是其他人吗？ （3）在底物结合和催化过程中，ARNT酶如何变化？ （4）路径外观中的其他跨膜酶如何完成 功能？ （5）途径中的其他跨膜酶如何容纳脂质糖载体 不依赖磷酸盐？为了解决这些问题，PI汇总了一项全面的研究计划 利用最先进的方法。此外，该计划包括一些针对的技术开发 在研究计划的背景下扩展电子显微镜的功能，并 该计划完成后，可能适用于其他研究计划。提出的研究 K99阶段将在很大程度上发生在哥伦比亚大学的著名环境中，该大学拥有充满活力的 结构生物学社区。这种环境肯定可以促进PI成功完成 拟议的研究计划并实现了他将过渡到独立研究职业的目标。