Spin Labeling Insights into Hydration Effects on Macromolecular Flexibility and Function

本项目将促进对水化动力学在调节蛋白质柔性、构象取样和生物分子功能中的作用的理解。水的水合作用在生物学中是必不可少的，并且是生物技术中适当设计含有酶的工程材料的关键因素。工作范围包括建立一个奥弗豪泽动态核极化（ODNP）光谱仪，同时设计和构建针对生物和材料样品优化的谐振器，以便可以询问材料周围水合水的局部扩散动力学。深入了解水的水合作用在大分子的灵活性将揭示结合NMR，EPR和ODNP调查的目标系统。这些包括了解分子拥挤如何调节折叠和动力学在天然非结构化蛋白质，如何从酶的活性位点远端突变可能会影响水化，因此构象采样和动力学，以及如何在水的水合作用的变化耦合到跨膜信号的整合膜蛋白。更广泛的影响这项研究中的技术科学培训遍及博士后助理，研究生和本科生以及高中生。该项目专门针对招募代表性不足的高中生进入佛罗里达大学的夏季科学教学计划。每年夏天，三名当地高中教师将参加筛选和提名两名学生参加为期六周的沉浸式研究体验。这些活动将提高公众对科学研究过程的认识，使这些机会很少的社区了解科学研究过程。其他推广活动包括在本科生进入大学第一年时对他们进行为期三到四年的培训。这些努力与参与贝克曼学者和HHMI生命科学计划相结合。

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.