Spin Labeling Insights into Hydration Effects on Macromolecular Flexibility and Function

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Intellectual MeritThis project will advance the understanding of the role water hydration dynamics in modulating protein flexibility, conformational sampling and biomolecular function. Water hydration is essential in biology and is a critical factor for proper design of engineered materials containing enzymes in biotechnology. The scope of work involves building an Overhauser dynamic nuclear polarization (ODNP) spectrometer with concomitant design and construction of resonators optimized for biological and materials based samples, such that local diffusion dynamics of hydration water surrounding the materials can be interrogated. Insights into the role of water hydration in macromolecular flexibility will be revealed from combined NMR, EPR and ODNP investigations of targeted systems. These include understanding how molecular crowding modulates folding and dynamics in natively unstructured proteins, how mutations distal from active sites in enzymes may influence hydration and hence conformational sampling and dynamics, and how changes in water hydration are coupled to transmembrane signaling of integral membrane proteins. Broader ImpactThe technical scientific training in this research spreads across post-doctoral associates, graduate and undergraduate students as well as high school students. The project specifically targets recruiting underrepresented high school students into the University of Florida's Summer Science Teaching Program. Three local high school teachers will participate in screening and nominating two students each summer for this six week research immersion experience. These activities would increase public awareness of the scientific research process to communities where these types of opportunities are rare. Other outreach includes training undergraduate students when they enter college in their first year, over a three to four year time period. These efforts are combined with participation in the Beckman Scholar's and HHMI Science for Life programs.