Upgrade Advanced EPR/DEER Spectroscopy Facility

升级先进的 EPR/DEER 光谱设施

• 批准号: RTI-2020-00727
• 负责人: Ernst, Oliver
• 金额: $ 10.93万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=693840
• 关键词: Upgrade; Advanced; EPR; DEER; Spectroscopy

Electron paramagnetic resonance (EPR) and double electronelectron resonance (DEER) spectroscopy are important structural biology methods to obtain information on protein dynamics and protein conformations, which is complementary to cryo-electron microscopy (cryo-EM) and x-ray crystallography. Continuous wave (cw) EPR and DEER are also precise molecular rulers to measure distances between two spin labels attached at specific sites of proteins. This is especially important when proteins are small or have flexible domains and thus exist in multiple conformations which cannot be distinguished by cryo-EM and x-ray crystallography. The lab of the applicant houses Canada's only CFI-funded EPR/DEER facility which since its opening in 2014 contributed to 14 high impact publications. An upgrade of the facility with new software and electronics as well as components providing higher sensitivity will allow our team of 10 researchers to perform molecular studies on protein conformations in the areas of GPCR signaling, nucleoside transport, protein homeostasis, enzyme mechanism, substrate transport through membranes of Gram negative pathogens, and protein folding. It will support the basic mechanistic understanding of protein function and will contribute to an understanding of a wide variety of diseases including heart, brain, metabolomic and neuro-degenerative diseases as well as cancer and others. Further, the advanced EPR/DEER facility complements the capabilities of a basic EPR instrument in the NMR facility of the University of Toronto Chemistry Department which serves Toronto's natural sciences and engineering researchers. Our group of 8 applicants from Toronto and two from other cities need to perform EPR/DEER measurements on limited protein quantities which require improved sensitivity of the EPR/DEER equipment.

电子顺磁共振（EPR）和双电子共振（DEER）光谱是获得蛋白质动力学和蛋白质构象信息的重要结构生物学方法，是与冷冻电镜（cryo-EM）和x射线晶体学相辅相成的。连续波（cw） EPR和DEER也是精确的分子标尺，用于测量附着在蛋白质特定位点的两个自旋标签之间的距离。当蛋白质很小或具有灵活的结构域，从而以多种构象存在时，这一点尤其重要，这些构象无法通过低温电镜和x射线晶体学进行区分。申请人的实验室拥有加拿大唯一的cfi资助的EPR/DEER设施，自2014年开放以来，该设施已发表了14篇高影响力出版物。升级后的设备配备了新的软件和电子设备，以及提供更高灵敏度的组件，将使我们的10名研究人员团队能够在GPCR信号传导、核苷转运、蛋白质稳态、酶机制、革兰氏阴性病原体膜底物转运和蛋白质折叠等领域进行蛋白质构象的分子研究。它将支持对蛋白质功能的基本机制理解，并将有助于理解各种疾病，包括心脏，大脑，代谢组学和神经退行性疾病以及癌症和其他疾病。此外，先进的EPR/DEER设施补充了多伦多大学化学系核磁共振设施中基本EPR仪器的能力，该设施为多伦多自然科学和工程研究人员提供服务。我们来自多伦多的8名申请人和来自其他城市的2名申请人需要对有限的蛋白质量进行EPR/DEER测量，这需要提高EPR/DEER设备的灵敏度。