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.

原子分辨率下生物大分子的可视化为深入理解基本生物过程和生命规则提供了重要信息。传统上，X射线晶体学和核磁共振是唯一两种可以通过实验确定三维结构的工具。 ( 3D) 原子水平生物大分子的结构，称为单粒子冷冻电子显微镜的先进显微镜技术的仪器和方法的最新进展。 （冷冻电镜）是分子可视化的颠覆性技术，冷冻电镜技术已成为确定大分子复合物或膜蛋白 3D 结构的首选技术，这对两种传统结构技术都构成了巨大的挑战。该奖学金将支持 PI 在加州理工学院冷冻电镜中心发展冷冻电镜专业知识，并将冷冻电镜技术应用于原子水平的生物分子结构表征。冷冻电镜加入 PI 的结构工具箱不仅会激发 PI 内布拉斯加大学林肯分校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拥有 Big10 大学但缺乏专业知识的州或冷冻电镜仪器。在该奖学金的支持下，PI 及其小组成员将接受有关金属蛋白和组装体结构表征的全面培训冷冻电镜，从网格准备、图像收集到 3D 图构建和模型开发，该项目将重点关注分别参与细菌氧化还原应激反应和过渡金属稳态的两个互连金属蛋白家族的结构研究。 -从本次培训中获得的 EM 将推动 PI 团队成功实现短期和长期的研究目标。该提案产生的结构信息将填补基本结构原则的关键空白。控制金属蛋白对生物和非生物氧化还原应激和抗生素耐药性的细菌反应从长远来看，将冷冻电镜专业知识添加到 PI 的跨学科技能中将使 PI 更好地在金属蛋白的结构机制研究中发挥主导作用。从教育角度来看，该提案将为研究生和博士后提供使用冷冻电镜技术的直接培训机会。冷冻电镜将被纳入 PI 在 UNL 进行的教学和培训活动，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 的冷冻电镜专业知识将得到提高。实现新的研究探索，促进整个校园的新合作，并促进创新研究。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进行评估，被认为值得支持标准。