Spin Labeling Insights into Hydration Effects on Macromolecular Flexibility and Function

自旋标记洞察水合对大分子灵活性和功能的影响

• 批准号: 1329467
• 负责人: Gail Fanucci
• 金额: $ 51.8万
• 依托单位: University of Florida
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1329467&HistoricalAwards=false
• 关键词: Spin; Labeling; Insights; into; Hydration

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.

智力价值这个项目将促进对水的水化动力学在调节蛋白质灵活性、构象采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，也是生物技术中含有酶的工程材料正确设计的关键因素。工作范围包括构建Overhauser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，以便可以询问材料周围水化水的局部扩散动力学。对目标体系的核磁共振、电子顺磁共振和ODNP的联合研究将揭示水的水化作用在大分子柔性中的作用。这些包括了解分子拥挤如何调节天然非结构蛋白的折叠和动力学，酶活性部位远端的突变如何影响水合作用从而影响构象采样和动力学，以及水合作用的变化如何与完整膜蛋白的跨膜信号相耦合。更广泛的影响这项研究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。该项目专门针对招收人数不足的高中生参加佛罗里达大学的暑期科学教学计划。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这次为期六周的研究沉浸体验。这些活动将提高公众对此类机会很少的社区的科研进程的认识。其他外展包括在本科生第一年进入大学时对他们进行培训，为期三到四年。这些努力与贝克曼学者和HHMI生命科学计划的参与相结合。