Fast protein liquid chromatography (FPLC) based ultra-pure protein preperations for the study of structural biology

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Ultra-pure protein and lipid mimetic preparations represent a gateway to higher level structural biology and biophysics approaches that range from solution and solid-state Nuclear Magnetic Resonance to cryo-electron microscopy, crystallography, and optical spectroscopy. The information gained by such approaches increases our fundamental knowledge of protein structure and furthers our understanding of how the changes proteins undergo in different cellular environments beget their function. The applicants and major users are members of the Biophysics Interdepartmental Group (BIG) at the University of Guelph, a multi-college initiative that promotes novel multidisciplinary research and training across the wide spectrum of biological and physical disciplines. This research group urgently requires a Fast Protein Liquid Chromatography (FPLS) system to replace an older unit that is currently end of life and to increase our protein biochemistry and protein structure-function research capacities. The requested system will support multiple leading-edge research programs that are delivering novel mechanistic insights into 1) strain specific conformational self-templating of intrinsically disordered proteins, 2) membrane protein function and folding, and 3) bacterial cell division and biofilm assembly. The requested investment in chromatography infrastructure is critical to our group's ability to remain at the forefront of protein biochemistry and structural biology research and training. The unit will support the research and training of >50 HQP at all levels in the applicants' and major users' labs and will further benefit the wider BIG community which has an annual intake of >25 graduate students. Critically, there is no similar equipment at the University of Guelph that has the available capacity to support the needs of all BIG investigators and their HQP. Thus, a delay in acquiring a fully functional and updated FPLC would significantly impact the success of our research network, hampering our workflows and disrupting HQP training and career progression. The requested system will be housed in the Advanced Analysis Center (AAC) at the University of Guelph, where it will be professionally maintained and fully accessible to the members of the BIG research group and others. In addition to HQP that will be trained from BIG labs, housing this technology in the AAC will greatly enhance equitable access and further the research capacity and output of scientists from across the University of Guelph, other universities in southern Ontario, and industrial scientists who regularly collaborate with University of Guelph researchers and access the AAC for scientific application and HQP training (>200 individual research groups access the AAC annually).