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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要抗生素耐药性细菌感染对人类健康构成巨大且日益严重的威胁。有一个迫切需要表征新的抗菌靶点并开发能够发现和利用的方法这些目标的抑制剂的表征。这里描述的研究重点是了解晚期围绕细菌细胞的肽聚糖细胞壁的组装步骤。细胞壁对于细菌的存活，使其成为抗生素的杰出目标。事实上，一些最重要的抗生素历史上，β-内酰胺和糖肽针对肽聚糖组装的后期步骤。从历史上看，由于催化这些后期步骤的酶的性质，研究起来极其困难。所需的底物、形成的产物以及许多肽聚糖生物合成的事实酶是多位膜蛋白或仅在与膜蛋白复合时才起作用。最近的我们实验室的技术创新推动了肽聚糖生物合成领域的发展相当大，我们能够追求以前从未研究过的具有挑战性的目标。这个项目有四个目标。目标 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。目标 2 侧重于了解 MurJ，即输出肽聚糖前体脂质 II 从细胞质到细胞表面，在细胞表面聚合产生肽聚糖。目标 3 侧重于表征 FtsW，这是唯一普遍保守的肽聚糖细菌中的聚合酶。目标 4 重点关注两种新型金黄色葡萄球菌细胞壁水解酶复合物，在未交联的肽聚糖整合到细胞壁之前对其进行处理。这四个目标将提供有关细菌细胞壁如何构建的基本信息。这些研究还将提供新的化学工具，检测和科学知识能够发现可用于治疗耐药性的抑制剂细菌感染。