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

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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.