Spin Labeling Insights into Hydration Effects on Macromolecular Flexibility and Function

本项目将促进对水合动力学在调节蛋白质柔韧性、构象采样和生物分子功能方面的作用的理解。水的水合作用在生物学中是必不可少的，也是生物技术中合理设计含酶工程材料的关键因素。工作范围包括建造一个奥弗豪瑟动态核极化（ODNP）光谱仪，同时设计和建造针对生物和基于材料的样品优化的谐振器，这样就可以询问材料周围水化水的局部扩散动力学。通过对目标体系的核磁共振、EPR和ODNP联合研究，将揭示水水合作用在大分子柔韧性中的作用。这些包括了解分子拥挤如何调节天然非结构蛋白质的折叠和动力学，远离酶活性位点的突变如何影响水合作用，从而影响构象采样和动力学，以及水水合作用的变化如何与整体膜蛋白的跨膜信号耦合。更广泛的影响本研究的技术科学培训遍及博士后、研究生和本科生以及高中生。该项目专门针对佛罗里达大学夏季科学教学计划招募代表性不足的高中生。每年夏天，三名当地高中教师将参与筛选和提名两名学生，参加为期六周的浸入式研究体验。这些活动将提高公众对这类机会很少的社区的科学研究过程的认识。其他拓展活动包括在本科学生进入大学一年级时对他们进行三到四年的培训。这些努力与贝克曼学者和HHMI生命科学项目的参与相结合。

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.