3 D Electron Microscopy of Macromolecules

大分子的 3D 电子显微镜

• 批准号: 7883922
• 负责人: Wah Chiu
• 金额: $ 89.29万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-13 至 2012-08-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7883922
• 关键词: Electron; Microscopy; Macromolecules

The NCMI is dedicated to the advancement of electron cryo-microscopy (cryo-EM) methodology for structure determination of nanomachines in their various functional states at the highest possible resolutions. Having established four state of the art electron cryomicroscopes and CCD cameras and accomplished de novo backbone traces of proteins in both viruses and molecular chaperonins, our Center is well poised to continue our research mission, including 4 core projects, 12 collaborative projects, 14 service projects, workshops, dissemination, training and maintaining an advisory board. In the core projects, we will 1) extend the resolution of single particle reconstruction beyond 4 A; 2) explore Zernike phase contrast for biological cryo- EM at subnanometer resolution; 3) develop techniques for "single particle" averaging in electron cryotomography at maximum possible resolution; 4) extend our data management infrastructure to fully document large scale cryo-EM datasets and processing. Specific aim 1 will bring a new level of capabilities to cryo-EM, providing molecular structures with a level of detail comparable to x-ray crystallography. The second aim will demonstrate the potentials and limitations of imaging with a theoretically promising new type of electron optics. Our third aim is targeted at uncovering unprecedented structure details about dynamic nanomachines operating in a native or near-native environment. The fourth aim will allow us to mine cryo-EM data easily to improve current methodologies and data sharing. The collaborative and service projects will cover a broad spectrum of specimens including viruses, molecular chaperonins, RNA, membrane and membrane associated proteins, and macromolecular assemblies associated with or inside cells. We will undertake a new effort in identifying projects of biomedical and translational relevance, through interactions with clinical and translational investigators. We will disseminate our software, experimental and computational protocols via workshops and web based materials. We will continue our tradition of training both local and distant students, postdocs and new investigators. We will maintain a dynamic membership of our advisory board. These efforts will extend cryo-EM's capabilities both at very high resolution at the interface with x-ray crystallography and at lower resolutions, relating macromolecular structures in the cell or cell-like environment.

NCMI致力于推进电子冷冻显微镜(Cryo-EM)结构方法学的发展 以尽可能高的分辨率测定各种功能状态下的纳米机器。拥有 建立了四台最先进的电子冷冻显微镜和ccd摄像机，完成了新的设计。 病毒和分子伴侣蛋白的骨架痕迹，我们的中心已经做好准备继续 我们的研究任务，包括4个核心项目，12个合作项目，14个服务项目，研讨会， 传播、培训和维持一个咨询委员会。在核心项目中，我们将1)延长 4 A以上的单粒子重建分辨率；2)探索用于生物低温的Zernike位相衬度。 3)发展电子冷冻层析成像中的“单粒子”平均技术 以最大可能的分辨率；4)将我们的数据管理基础架构扩展到完全 记录大规模冷冻-EM数据集和处理。特定目标1将带来新的能力水平 到冷冻-EM，提供了可与X射线结晶学相媲美的详细分子结构。这个 第二个目标将用一种理论上有前景的新型成像来展示成像的潜力和局限性 电子光学的。我们的第三个目标是揭示关于Dynamic的前所未有的结构细节 在原生或接近原生环境中运行的纳米机器。第四个目标将允许我们开采低温电磁 数据可轻松改进现有方法和数据共享。协作和服务项目将 涵盖广泛的样本，包括病毒、分子伴侣、RNA、膜和 膜相关蛋白，以及与细胞相关或在细胞内的大分子组件。我们会 通过互动，在确定与生物医学和翻译相关的项目方面做出新的努力 与临床和转化性研究人员合作。我们将传播我们的软件，试验性和 通过研讨会和基于网络的材料进行计算。我们将继续我们的训练传统 包括本地和远程学生、博士后和新的调查人员。我们将保持一个动态的成员资格 我们的顾问委员会。这些努力将扩展冷冻-EM的能力，在非常高的分辨率下 与X射线结晶学的接口，并在较低的分辨率下，关联细胞或 类似细胞的环境。