Electron Microscopy of Biological Macromolecules

生物大分子的电子显微镜

• 批准号: 9325991
• 负责人: Eva Nogales
• 金额: $ 7.9万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325991
• 关键词: Address; Algorithms; Automation; Bayesian Method; Biochemical; Biological; Cell physiology; Collection; Complex; Computers; Contract Services; Cost Sharing; Coupled; Cryoelectron Microscopy; Data; Data Collection; Data Set; Detection; Devices; Disease; Educational Status; Electron Microscope; Electron Microscopy; Electrons; Ensure; Environment; Equipment; Foundations; Future; Goals; Heterogeneity; Hour; Human Resources; Image; Image Analysis; Individual; Lead; Leadership; Macromolecular Complexes; Measures; Medical; Methodology; Molecular Machines; Motion; Output; Price; Proteins; Research; Research Infrastructure; Research Personnel; Research Project Grants; Resolution; Resources; Risk-Taking; Sampling; Series; Solid; Sorting - Cell Movement; Structure; Techniques; Therapeutic; X-Ray Crystallography; Yang; biological systems; computing resources; cost; detector; flexibility; image processing; image reconstruction; improved; innovation; insight; instrument; instrumentation; interest; macromolecular assembly; macromolecule; member; microscopic imaging; programs; public health relevance; scientific computing; structural biology; success

 DESCRIPTION (provided by applicant): Visualizing the structures of biological molecular machines is crucial for a mechanistic understanding of cellular function. Cryo-EM is an emergent structural biology technique that is ideally suited to study large macromolecular assemblies, even if they are only available in minute amounts, or if multiple states co-exist in th sample. The cryo-EM field has just entered a new era with the use of high-end electron microscopes coupled to new direct electron detection devices (DDDs) that has led to high resolution structural information, in some cases from only one or a few imaging sessions. As technical barriers open up and the field matures, the impact of new cryo-EM studies will more and more be defined by the output of biological insight. Such will be measured by the relevance of the biological system under study and by whether the experimental strategy is able to provide answers to key functional questions. This scenario requires the synergistic integration of cryo-EM and biochemical expertise in the context of studies that address fundamental biological questions. This program project brings together such set of complementary expertise and provides a mechanism to integrate them seamlessly to tackle biological studies of great medical significance, providing a supportive and coherent environment that promotes interactions among the different projects and advances each of them individually, as well as the cryo-EM field as a whole. This Program Project also provides a mechanism whereby the large costs of operating high-end electron microscopes, associated equipment and computational capabilities are effectively shared among a number of research projects that rely on such state-of-the-art instrumentation. As cryo-EM and image reconstruction are still developing rapidly, optimal use of this methodology requires rigorous retraining and is best facilitated by specialized support personnel and intensive discussion among members of a collaborative group as proposed in this program project. Our ultimate goal is to maintain and enhance Berkeley's contributions to the structural biology field. We will support and employ a physical and intellectual infrastructure that allows us to carry out state-of-the-art cryo-EM studies of essential macromolecular complexes and push the capabilities of the cryo-EM technique. This proposal capitalizes on (1) the tradition of innovation and risk-taking within our PPG and years of leadership and expertise in the field of biological electron microscopy and structural biology, (2) the implementation at Berkeley of recent technical breakthroughs in the cryo-EM field (e.g. DDDs, automation, Bayesian methods of image processing), (3) unique biochemical expertise in a large number of biological systems of outstanding biomedical relevance, and (4) the unique local computer resources at the National Energy Research Scientific Computing Center (NERSC), which is hosted at LBNL.

 描述(由申请人提供)：可视化生物分子机器的结构对于从机制上理解细胞功能至关重要。冷冻-EM是一种新兴的结构生物学技术，非常适合于研究大分子组件，即使它们只有少量可用，或者如果样品中共存多种状态。低温电磁场领域刚刚进入了一个新时代，高端电子显微镜的使用与新的直接电子探测设备(DDD)相结合，从而获得了高分辨率的结构信息，在某些情况下，只有一次或几次成像会议。随着技术壁垒的开放和该领域的成熟，新的低温电磁研究的影响将越来越多地由生物学洞察力的输出来定义。这将通过正在研究的生物系统的相关性以及实验战略是否能够为关键的功能问题提供答案来衡量。这一设想需要在解决基本生物学问题的研究背景下，协同整合低温EM和生化专业知识。该计划项目汇集了这些互补的专业知识，并提供了一种机制，将它们无缝地整合起来，以处理具有重大医学意义的生物学研究，提供一个支持性和一致性的环境，促进不同项目之间的互动，并单独推进每个项目，以及低温电磁领域作为一个整体。该计划项目还提供了一种机制，使操作高端电子显微镜、相关设备和计算能力的大量成本有效地在依赖这种最先进仪器的若干研究项目之间分摊。由于冷冻-EM和图像重建仍在快速发展，优化使用这一方法需要严格的再培训，最好是通过专业支持人员和本计划项目中提议的协作小组成员之间的深入讨论来促进这一方法。我们的最终目标是保持和提高伯克利在结构生物学领域的贡献。我们将支持和使用物质和智力基础设施 这使我们能够对重要的大分子络合物进行最先进的冷冻-EM研究，并推动低温-EM技术的能力。这项建议利用了(1)PPG的创新和冒险传统以及在生物电子显微镜和结构生物学领域的多年领导和专业知识，(2)在伯克利实施的冷冻-EM领域最近的技术突破(例如，DDDS、自动化、贝叶斯图像处理方法)，(3)在大量具有突出生物医学相关性的生物系统中的独特生化专业知识，以及(4)设在LBNL的国家能源研究科学计算中心(NERSC)独特的本地计算机资源。