Upgrade Advanced EPR/DEER Spectroscopy Facility

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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.