Upgrade Advanced EPR/DEER Spectroscopy Facility

升级先进的 EPR/DEER 光谱设施

基本信息

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

项目摘要

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）和双电子电子共振（DEER）波谱是获取蛋白质动力学和蛋白质构象信息的重要结构生物学方法，与冷冻电子显微镜（cryo-EM）和X射线晶体学互补。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上的两个自旋标签之间的距离。当蛋白质较小或具有灵活的结构域并因此存在无法通过冷冻电镜和 X 射线晶体学区分的多种构象时，这一点尤其重要。申请人的实验室拥有加拿大唯一一个由 CFI 资助的 EPR/DEER 设施，自 2014 年开放以来，已发表了 14 篇高影响力出版物。通过使用新软件和电子设备以及提供更高灵敏度的组件对设施进行升级，我们的 10 名研究人员团队将能够对 GPCR 信号传导、核苷转运、蛋白质稳态、酶机制、底物转运等领域的蛋白质构象进行分子研究。革兰氏阴性病原体的膜和蛋白质折叠。它将支持对蛋白质功能的基本机制理解，并将有助于理解多种疾病，包括心脏、大脑、代谢组和神经退行性疾病以及癌症等。此外，先进的 EPR/DEER 设施补充了多伦多大学化学系 NMR 设施中基本 EPR 仪器的功能，为多伦多的自然科学和工程研究人员提供服务。我们来自多伦多的 8 名申请者和来自其他城市的两名申请者需要对有限的蛋白质数量进行 EPR/DEER 测量，这需要提高 EPR/DEER 设备的灵敏度。