Discovery and characterization of new bacterial cell wall targets and inhibitors to treat resistant infections

治疗耐药感染的新细菌细胞壁靶点和抑制剂的发现和表征

• 批准号: 10078251
• 负责人: Daniel Kahne
• 金额: $ 76.72万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078251
• 关键词: Address; Anabolism; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Cell Separation; Cell Wall; Cell division; Cell membrane; Cell surface; Cells; Cellular biology; Chemicals; Collaborations; Collection; Complex; Cytoplasm; Development; Enzymatic Biochemistry; Enzymes; Family; Foundations; Glean; Glucosaminidase; Glycopeptides; Goals; Grant; Health; Human; Hydrolase; In Vitro; Infection; Knowledge; Lead; Learning; Letters; Light; Lipids; Membrane Potentials; Membrane Proteins; Methods; Molecular Conformation; Nature; Organism; Pathogenesis; Pathway interactions; Peptidoglycan; Peptidyltransferase; Physiology; Play; Polymerase; Polymers; Process; Proteins; Publishing; Recording of previous events; Research; Research Support; Resistance; Resistance to infection; Role; Staphylococcus aureus; Staphylococcus aureus infection; Structure; Vancomycin; Variant; amidase; analog; bacterial genetics; bacterial resistance; base; beta-Lactams; crosslink; daughter cell; design; enzyme structure; genetic approach; high throughput screening; inhibitor/antagonist; insight; muramyl-NAc-(pentapeptide)pyrophosphoryl-undecaprenol; mutant; novel; novel strategies; programs; protein complex; reconstitution; response; technological innovation; tool

Project Summary Antibiotic-resistant bacterial infections pose a substantial and growing threat to human health. There is a pressing need to characterize new antibacterial targets and to develop methods that enable discovery and characterization of inhibitors for these targets. The research described here focuses on understanding late steps in the assembly of the peptidoglycan cell wall that surrounds bacterial cells. The cell wall is essential for bacterial survival, making it an outstanding target for antibiotics. Indeed, some of the most important antibiotics in history, the beta-lactams and the glycopeptides, target late steps in peptidoglycan assembly. Historically, the enzymes that catalyze these late steps have been extremely difficult to study due to the nature of the substrates that are required, the products that are formed, and the fact that many peptidoglycan biosynthetic enzymes are polytopic membrane proteins or function only when complexed to a membrane protein. Recent technological innovations made in our labs have advanced the field of peptidoglycan biosynthesis considerably, and we are able to pursue challenging targets that have not previously been studied. This project has four aims. Aim 1 involves further development of chemical tools to enable mechanistic and structural studies of cell wall biosynthetic enzymes. Aim 2 focuses on understanding MurJ, the flippase that exports the peptidoglycan precursor Lipid II from the cytoplasm to the cell surface where it is polymerized to produce peptidoglycan. Aim 3 focuses on characterizing FtsW, the only universally conserved peptidoglycan polymerase in bacteria. Aim 4 focuses on two novel Staphylococcus aureus cell wall hydrolase complexes that process uncrosslinked peptidoglycan before it is integrated into the cell wall. These four aims will provide fundamental information on how the bacterial cell wall is built. The studies will also provide new chemical tools, assays, and scientific knowledge to enable the discovery of inhibitors that may be useful for treating resistant bacterial infections.

项目摘要 抗抗生素的细菌感染对人类健康构成了巨大的威胁。有一个 迫切需要表征新的抗菌靶标，并开发出启用发现和的方法 这些靶标的抑制剂的表征。这里描述的研究重点是理解迟到 围绕细菌细胞的肽聚糖细胞壁组装的步骤。细胞壁对于 细菌生存，使其成为抗生素的杰出靶标。确实，一些最重要的抗生素 在历史上，β-内酰胺和糖肽，靶向肽聚糖组装的后期步骤。从历史上看， 催化这些后期步骤的酶由于性质而难以研究 所需的底物，形成的产品以及许多肽聚糖生物合成的事实 酶是多重膜蛋白或仅在复合到膜蛋白时功能。最近的 在我们的实验室中进行的技术创新已经提高了肽聚糖生物合成领域 相当多的是，我们能够追求以前没有研究过的具有挑战性的目标。这个项目 有四个目标。 AIM 1涉及进一步开发化学工具以实现机械和结构 细胞壁生物合成酶的研究。 AIM 2专注于理解Murj，这是出口的Flippase 肽聚糖前体脂质II从细胞质到细胞表面聚合以产生 肽聚糖。 AIM 3专注于表征FTSW，这是唯一普遍保守的肽聚糖 细菌中的聚合酶。 AIM 4专注于两种新型的金黄色葡萄球菌细胞壁水解酶配合物， 在将其整合到细胞壁中之前，过程无链接肽聚糖。这四个目标将提供 有关细菌细胞壁的建造方式的基本信息。研究还将提供新的化学工具 测定和科学知识，以发现可能有助于治疗抗性的抑制剂 细菌感染。