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制剂代表了通向更高水平的结构生物学和生物物理学方法的大门，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。通过这些方法获得的信息增加了我们对蛋白质结构的基础知识，并进一步加深了我们对蛋白质在不同细胞环境中经历的变化如何产生其功能的理解。 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，旨在促进跨广泛的生物和物理学科的新颖的多学科研究和培训。该研究小组迫切需要快速蛋白质液相色谱 (FPLS) 系统来替换目前已报废的旧装置，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。所请求的系统将支持多个前沿研究项目，这些项目为 1) 本质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提供新颖的机制见解。 所要求的色谱基础设施投资对于我们集团保持在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各级超过 50 个 HQP 的研究和培训，并将进一步使每年招收超过 25 名研究生的更广泛的 BIG 社区受益。至关重要的是，圭尔夫大学没有类似的设备能够满足所有 BIG 研究人员及其总部的需求。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。 所请求的系统将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维护，并可供 BIG 研究小组成员和其他人完全访问。除了将在 BIG 实验室进行培训的 HQP 之外，将这项技术纳入 AAC 将极大地提高公平获取机会，并进一步提高来自圭尔夫大学、安大略省南部其他大学的科学家以及定期参与的工业科学家的研究能力和产出。与圭尔夫大学研究人员合作，并访问 AAC 进行科学应用和 HQP 培训（每年有超过 200 个独立研究小组访问 AAC）。