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

项目总结/摘要 为响应NOT-GM-22-017，关于以下事项的行政补充材料可用性的通知 为NIGMS获奖者购买设备，我们要求资金购买一台纯的ÄKTA 25 M液相色谱系统和一套最适合于 在母基金R 01 GM 139034中提出的工作。家长补助金旨在建立 病毒识别其宿主细胞的机制，经历构象变化， 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组分在 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中渗透。单独的病毒体成分，病毒宿主- 附着细胞器，完整的噬菌体颗粒和宿主细胞的各种组分-寡核苷酸， 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、低温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方法的成功至关重要。 问题研究这种制备可以通过<$KTA纯25 M液相色谱法进行 该系统配备有所提出的一组色谱柱， 容量/高分辨率阴离子交换柱，分析型（小容量和高分辨率） 尺寸排阻柱和制备型（大体积）尺寸排阻柱，其中后两种 针对不同分子量的大分子的类型。净化系统和 列将提供给其他NIH资助的研究者，如果需要的话。

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.