Atomic-level probing of the peptidoglycan biosynthetic machinery in bacterial cell wall biogenesis

细菌细胞壁生物发生中肽聚糖生物合成机制的原子水平探测

• 批准号: 10685947
• 负责人: Allison H Williams
• 金额: $ 41.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685947
• 关键词: Acetylesterase; Affinity Chromatography; Antibiotics; Bacteria; Bacterial Physiology; Binding; Binding Proteins; Biochemical; Biochemical Reaction; Biogenesis; Biological; Biological Assay; Biophysics; Cell Wall; Cell division; Cells; Communication; Complement; Complex; Coupled; Cryoelectron Microscopy; Data; Development; Drug resistance; Enzymes; Future; Genetic; Goals; Gram-Negative Bacteria; Hot Spot; Hydrolysis; Infection; Intracellular Transport; Lytic; Maps; Mass Spectrum Analysis; Membrane; Metabolic; Metabolism; Microbe; Modification; Molecular; Molecular Conformation; Multienzyme Complexes; Nature; Neisseria; Neisseria gonorrhoeae; Neisseria meningitidis; Organelles; Penicillin-Binding Proteins; Peptidoglycan; Peptidoglycan glycosyltransferase; Pilum; Polymerase; Polysaccharides; Positioning Attribute; Process; Productivity; Protein-Protein Interaction Map; Proteins; Public Health; Resistance; Resolution; Role; Site; Structure; Testing; Time; Validation; Visualization; Work; X-Ray Crystallography; analog; antimicrobial; biochemical tools; cell envelope; cell motility; drug development; fighting; human pathogen; insight; macromolecule; member; nanobodies; nanomachine; new therapeutic target; next generation; particle; pathogen; pathogenic bacteria; protein complex; structural biology; tool

The emergence of microbes resistant to even the most powerful antibiotics represents a serious threat to global public health. The coordinated action of the bacterial machinery of peptidoglycan (PG) synthesis, a process essential for bacterial viability, represents an obvious target for the development of new antibiotics. In this proposed project, we aim to define the molecular interactions of the components of the PG degradation apparatus, consisting of enzymes involved in glycan chain hydrolysis or modification. Our previous studies in Neisseria meningitidis showed that targeting a hot spot on a single lytic transglycosylase (LgtA) also disables the function of the PG-modifying enzyme, Ape1, leading to a disruption of PG assembly, and results in an aberrant peptidoglycan composition, making this pathogen unable to survive in the host. Our studies will reveal, in molecular detail, how these peptidoglycan degrading enzymes work in concert to assemble the bacterial cell wall. Specifically, we will define, in a comprehensive way, how a network of lytic transglycosylases (LtgA, LtgD, LtgE) and their protein binding partners work to facilitate peptidoglycan degradation and the insertion of organelles into the bacterial cell envelope. In this project, we will utilize biochemical and biological approaches to probe protein-protein and enzyme-substate interactions of the various lytic transglycosylases, combined with determination of the molecular basis of activity of the multienzyme complexes in PG metabolism. Genetic modifications of the components of the PG biosynthetic nanomachine will be used to test the observations from our structural studies. Our approach, utilizing high resolution x-ray crystallographic tools along with cryo-EM single particle analysis, will allow visualization of the action of enzymes in PG assembly and degradation and should provide mechanistic insights into their orchestrated activity during the insertion of new PG during cell wall assembly and bacterial cell division. Our studies will lead the way towards the development of new therapies targeting multiple peptidoglycan metabolic enzymes.

即使是最强大的抗生素也有耐药性的微生物的出现对人类构成了严重威胁