Structural-functional characterization of a hyperactive antifreeze protein

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With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding the research of Dr. Krisztina Varga and Dr. Daniel Levy from the University of Wyoming to structurally and functionally characterize a hyperactive insect antifreeze protein. Antifreeze proteins are produced in several organisms that live in cold climates and contribute to their resistance to freezing. They bind irreversibly to the surface of ice crystals and inhibit their growth, however, the exact mechanism at the molecular level is not fully understood. The investigators will determine the structure of the antifreeze protein, examine its water binding properties, and investigate its mechanism of freeze inhibition in live cells. The results of this research will provide a better fundamental understanding of molecular processes characteristic of antifreeze proteins interacting with ice and expand their practical applications, for example, in protecting foods from damage by freezing. The project will have a broader impact by providing opportunities for underrepresented groups of graduate and undergraduate students to participate in scientific research, many of them being first generation university students from rural areas. The project will be integrated into outreach programs for community college, high school and grade school students.This research will probe the structure and mechanism of action of the hyperactive insect antifreeze protein ApAFP752. The research will provide a better understanding of how antifreeze proteins interact with ice and provide cryoprotection to organisms that live in cold environments. The approaches are comprehensive and combine biochemistry, NMR spectroscopy, and confocal microscopy for both in vitro and in vivo studies and will utilize the synergy of the expertise of the investigators in these areas. This research project will be undertaken to (1) structurally characterize the antifreeze protein by NMR spectroscopy, (2) explore its water binding properties and interactions (e.g. hydrogen bonding), and (3) investigate its cryoprotective mechanism in Xenopus live cells. The comprehensive approach will provide unique insights into the underlying molecular mechanism of cryopreservation.This project is co-funded by the Experimental Program for the Stimulation of Competitive Research (EPSCoR).