Structural and functional determinants of decision-making in bacteriophage host recognition

噬菌体宿主识别决策的结构和功能决定因素

• 批准号: 10798641
• 负责人: Petr G Leiman
• 金额: $ 14.27万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-10 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10798641
• 关键词: Acceleration; Administrative Supplement; Anions; Antibiotic Therapy; Bacteriophages; Biological Models; Biophysics; Cell surface; Cells; Column Chromatography; Complex; Cryoelectron Microscopy; Decision Making; Diagnostic; Exclusion; Funding; High Pressure Liquid Chromatography; Individual; Liquid Chromatography; Membrane Proteins; Modeling; Molecular Conformation; Molecular Weight; National Institute of General Medical Sciences; O Antigens; Oligonucleotides; Organelles; Penetration; Polysaccharides; Preparation; Process; Research Personnel; Resolution; Role; Structure; System; Therapeutic; United States National Institutes of Health; Viral; Virion; Virus; Work; X-Ray Crystallography; antibiotic resistant infections; cell envelope; equipment acquisition; macromolecule; parent grant; particle; public health relevance; response; success; tool; transforming virus

PROJECT SUMMARY/ABSTRACT In response to NOT-GM-22-017, Notice of Availability of Administrative Supplements for Equipment Purchases for NIGMS Awardees, we request the funds to purchase one ÄKTA pure 25 M liquid chromatography system and a set of chromatography columns most suitable for the work proposed in the parent grant R01GM139034. The parent grant aims to establish the mechanisms by which viruses recognize their host cells, undergo a conformational change, and irreversibly attach to the cell surface. Using several bacterial viruses (bacteriophages) as model systems, the project aims to determine the role of different phage virion components in the attachment process, the energetics of host cell surface recognition, and the structure of the phage particle in the midst of host cell envelope penetration. Individual virion components, viral host- attachment organelles, complete phage particles, and various components of the host cell – oligo- and polysaccharides, including O-antigens, and membrane proteins – will be studied using X-ray crystallography, cryo-electron microscopy, and solution biophysics. High quality and high purity preparations of macromolecules and their complexes are essential for the success of these studies. Such preparations can be made possible by the ÄKTA pure 25 M liquid chromatography system equipped with the proposed set of chromatography columns that include a high capacity/high resolution anion exchange column, analytical (small volume and high resolution) size exclusion columns and preparative (large volume) size exclusion columns with the latter two types targeting different molecular weights of macromolecules. The purification system and the columns will be made available to other NIH-funded investigators should the need arise.

项目概要/摘要 为了回应 NOT-GM-22-017，行政补充的可用性通知 为 NIGMS 获奖者购买设备，我们请求资金购买一台 AKTA pure 25M液相色谱系统及一套最适合的色谱柱 母基金 R01GM139034 中提议的工作。家长补助金旨在建立 病毒识别宿主细胞、经历构象变化的机制，以及 不可逆地附着在细胞表面。使用几种细菌病毒（噬菌体）作为模型 系统，该项目旨在确定不同噬菌体病毒体成分在 附着过程、宿主细胞表面识别的能量学以及噬菌体的结构 颗粒在宿主细胞包膜穿透过程中。单个病毒体成分，病毒宿主 附着细胞器、完整噬菌体颗粒和宿主细胞的各种成分 – 寡聚- 和多糖，包括 O 抗原和膜蛋白 - 将使用 X 射线进行研究 晶体学、冷冻电子显微镜和溶液生物物理学。高品质、高纯度 大分子及其复合物的制备对于这些成功至关重要 研究。此类制备可以通过 äKTA pure 25 M 液相色谱来实现 系统配备了建议的一套色谱柱，其中包括高 大容量/高分辨率阴离子交换柱，分析型（小体积和高分辨率） 尺寸排阻色谱柱和制备型（大体积）尺寸排阻色谱柱，后两者 针对不同分子量的大分子的类型。净化系统和 如果需要，列将提供给其他 NIH 资助的研究人员。

项目成果

Quantitative description of a contractile macromolecular machine.

• DOI: 10.1126/sciadv.abf9601
• 发表时间: 2021-06
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Fraser A;Prokhorov NS;Jiao F;Pettitt BM;Scheuring S;Leiman PG
• 通讯作者: Leiman PG

Reprogramming bacteriophage host range: design principles and strategies for engineering receptor binding proteins.

• DOI: 10.1016/j.copbio.2021.02.006
• 发表时间: 2021-04
• 期刊: Current opinion in biotechnology
• 影响因子: 7.7