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

基于快速蛋白质液相色谱 (FPLC) 的超纯蛋白质制剂，用于结构生物学研究

• 批准号: RTI-2022-00047
• 负责人: Ryan, Scott
• 金额: $ 9.62万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=742918
• 关键词: Fast; protein; liquid; chromatography; FPLC

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).

超纯蛋白质和脂质模拟物制剂代表了通往更高水平的结构生物学和生物物理学方法的途径，这些方法的范围从溶液和固态核磁共振到冷冻电子显微镜，晶体学和光谱学。通过这种方法获得的信息增加了我们对蛋白质结构的基本知识，并进一步了解蛋白质在不同细胞环境中如何发生变化以产生其功能。 申请人和主要用户是圭尔夫大学生物物理学跨部门小组（BIG）的成员，这是一个多学院倡议，旨在促进跨生物和物理学科的广泛领域的新的多学科研究和培训。该研究小组迫切需要一个快速蛋白质液相色谱（FPLS）系统，以取代目前寿命结束的旧设备，并提高我们的蛋白质生物化学和蛋白质结构-功能研究能力。所要求的系统将支持多个前沿研究项目，这些项目将提供新的机制见解，包括1）内在无序蛋白质的菌株特异性构象自模板，2）膜蛋白功能和折叠，以及3）细菌细胞分裂和生物膜组装。 所要求的色谱基础设施投资对于我们集团保持蛋白质生物化学和结构生物学研究和培训前沿的能力至关重要。该单位将支持申请人和主要用户实验室中所有级别的>50名HQP的研究和培训，并将进一步使每年招收>25名研究生的更广泛的BIG社区受益。至关重要的是，在圭尔夫大学没有类似的设备，有能力支持所有BIG调查人员及其HQP的需求。因此，延迟获得功能齐全且更新的FPLC将严重影响我们研究网络的成功，阻碍我们的工作流程并扰乱HQP培训和职业发展。 所要求的系统将被安置在圭尔夫大学的高级分析中心（AAC），在那里它将得到专业的维护，并完全供BIG研究小组的成员和其他人使用。除了将从BIG实验室接受培训的HQP外，将这项技术安置在AAC中将大大提高公平获取，并进一步提高来自圭尔夫大学、安大略南部其他大学的科学家的研究能力和产出，和工业科学家，他们定期与圭尔夫大学的研究人员合作，并访问AAC进行科学应用和HQP培训（每年有超过200个研究小组访问AAC）。