Pressure apparatus for nuclear magnetic resonance studies of protein excited states

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We request funds to be able to perform Nuclear Magnetic Resonance (NMR) studies at pressures between 1 bar and 2000 bar. The one-of-a-kind apparatus at the University of Toronto would be installed in an existing protein NMR facility and used by various researchers. Temperature and pressure are critically useful parameters in structure studies of macromolecules and proteins. Our main research interests include photo-switchable proteins and membrane proteins. In both cases, we may describe the proteins in terms of a ground state and one or more low-lying excited states. The excited states and excited state intermediates are often associated with the active form of the protein and are often the most elusive to understand. The application of a technique called NMR dispersion spectroscopy allows us to tease out the energy landscape associated with the protein and its excited state(s). By repeating these measurements as a function of pressure we can learn fundamental information about how the excited states are constructed. The apparatus permits us to make great leaps in our current research (e.g. specific volumes and compressibilities of excited states, water penetration of folded proteins, new information on photoswitchable proteins) and study pressure-dependent phenomena by high resolution NMR. We also believe that that the apparatus will forge new collaborations and establish new research niches for the applicants.