RII Track-4: Molecular Visualization by Single Particle Cryo-Electron Microscopy

RII Track-4：单粒子冷冻电子显微镜的分子可视化

• 批准号: 2033441
• 负责人: Limei Zhang
• 金额: $ 21.58万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2033441&HistoricalAwards=false
• 关键词: RII; Track; Molecular; Visualization; Single

Visualization of biomacromolecules at atomic resolutions provides important informationfor the in-depth understanding of the fundamental biological processes and the rules of life.Traditionally, X-ray crystallography and nuclear magnetic resonance have been the only twoavailable structural tools that can experimentally determine three-dimensional (3D) structures ofbiomacromolecules at atomic levels. Recent advances in the instrumentation and methodology ofthe advanced microscopy technique called single-particle cryo-electron microscopy (cryo-EM) isa game-changer for molecular visualization. The cryo-EM technique has become the preferredtechnique for determining the 3D structure of large macromolecular complex or membraneproteins that have been dauntingly challenging to either of the two conventional structuraltechniques. This fellowship will support the PI to develop the cryo-EM expertise at the Cryo-EMCenter of the California Institute of Technology and apply the cryo-EM technique to the structuralcharacterization of biomolecules at the atomic level. Adding cryo-EM to the PI’s structural toolboxwill not only stimulate new research directions in the PI’s research group at the University ofNebraska-Lincoln, but also benefit many research groups across the state of Nebraska, which isthe only state that hosts a Big10 university but without expertise or instrumentation for cryo-EM.With the support of this fellowship, the PI and her group member will receivecomprehensive training on structural characterization of metalloproteins and assemblies by cryo-EM, from grid preparation, image collection to 3D map construction and model development. Inparticular, this project will focus on the structural investigation of two families of interconnectedmetalloproteins involved in bacterial redox stress response and transition metal homeostasis,respectively. The expertise in cryo-EM acquired from this training will propel the PI’s group tosuccessfully achieve research goals in both the short and long term. The structural informationresulted from the proposal will fill critical gaps in fundamental structural principles that governthe metalloproteins in the bacterial response to biotic and abiotic redox stresses and antibioticresistance. In the long run, adding the cryo-EM expertise to the PI’s interdisciplinary skillset willbetter position the PI to play a leading role in structure-mechanism investigations ofmetalloproteins in redox biology. From an educational aspect, this proposal will provide directtraining opportunities to graduate students and postdocs using the cryo-EM technique. In addition,the acquired knowledge of the application of cryo-EM will be integrated into the teaching andtraining activities conducted by the PI at UNL, aligning with the educational goal in the PI’s NSFCAREER award. Furthermore, by working with the colleagues at the University of Nebraskasystem, the PI’s cryo-EM expertise will enable new research explorations and catalyze newcollaboration across the campus and promote innovation research.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

生物大分子在原子水平上的可视化研究为深入理解基本的生物过程和生命规律提供了重要的信息，传统上，X射线晶体学和核磁共振是仅有的两种可以在原子水平上实验确定生物大分子三维结构的结构工具.单粒子低温电子显微镜（cryo-EM）是一种先进的显微镜技术，其仪器和方法学的最新进展伊萨了分子可视化的游戏规则。cryo-EM技术已成为确定大分子复合物或膜蛋白三维结构的首选技术，这对两种传统结构技术中的任何一种都具有巨大的挑战性。该奖学金将支持PI在加州理工学院的Cryo-EM中心发展cryo-EM专业知识，并将cryo-EM技术应用于原子水平的生物分子结构表征。将cryo-EM添加到PI的结构工具箱中不仅会刺激内布拉斯加大学林肯分校PI研究小组的新研究方向，而且还会使内布拉斯加州的许多研究小组受益，内布拉斯加州是唯一一个拥有十大大学但没有cryo-EM专业知识或仪器的州。主要研究者和她的小组成员将接受关于冷冻电镜金属蛋白和组装体结构表征的综合培训，从网格准备，图像收集到3D图构建和模型开发。特别是，这个项目将集中在两个家庭的interconnectedmetalloproteins参与细菌氧化还原应激反应和过渡金属稳态的结构调查，分别。从这次培训中获得的cryo-EM专业知识将推动PI的团队成功实现短期和长期的研究目标。从该提案中获得的结构信息将填补基本结构原则的关键空白，这些原则决定了细菌对生物和非生物氧化还原应激和耐药性的反应中的金属蛋白。从长远来看，将cryo-EM专业知识添加到PI的跨学科技能中将更好地定位PI，使其在氧化还原生物学中金属蛋白的结构机制研究中发挥主导作用。从教育的角度来看，这项建议将提供直接培训的机会，研究生和博士后使用冷冻EM技术。此外，所获得的cryo-EM应用知识将被整合到由PI在UNL进行的教学和培训活动中，与PI的NSFCAREER奖的教育目标保持一致。此外，通过与内布拉斯加大学系统的同事合作，PI的cryo-EM专业知识将使新的研究探索成为可能，并促进整个校园的新合作，促进创新研究。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。