An ODNP Probe to Study Hydration Dynamics in Membrane Protein

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 DESCRIPTION (provided by applicant): The proposed research focuses on the development of a cost-effect, turn-key probe for liquid-state Overhauser Dynamic Nuclear Polarization (ODNP) spectroscopy to study the site-specific translational dynamics of water molecules located at the interface of bio-macromolecules such as membrane proteins. It will allow researchers to readily perform ODNP experiments in a state-of-the-art commercially available X-Band electron paramagnetic resonance (EPR) spectrometer. In recent years, DNP has proven to be a robust method to increase signal intensities in NMR experiments in laboratories around the world and substantial progress has been made in adapting DNP for solid- and solution-state NMR spectroscopy. This progress has sparked a new interest in ODNP spectroscopy. Although the method is known since the 1960s it has just recently been applied successfully to study the site- specific translational dynamics of water located at the interface o large bio-macromolecules such as membrane proteins. ODNP can map out the local and site-specific hydration dynamics landscape of membrane proteins and lipid membranes and can provide critical information about the protein structure and dynamics. Currently there is no commercial probe available to readily start 9 GHz ODNP experiments; which means the method is only available to a small number of labs that are tech savvy and willing to tinker with their instrumentation. In this SBIR Phase I application we propose to develop a turn-key, cost-effective ODNP probe to be used in a state-of-the-art continuous wave X-Band EPR spectrometer, which can be found in many academic spectroscopy facilities. The successful development of this technology will provide researchers access to incorporate ODNP spectroscopy in their research routine without the hassle of troubleshooting home-built equipment. This will greatly proliferate the method and is of large interest to many projects funded by the U.S. National Institutes of Health.