US-Switzerland Collaborative Research on Cryo-Electron Microscopy

美国-瑞士冷冻电子显微镜合作研究

• 批准号: 8900543
• 负责人: Edward Egelman
• 金额: $ 0.82万
• 依托单位: Yale University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-15 至 1992-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8900543&HistoricalAwards=false
• 关键词: US; Switzerland; Collaborative; Research; Cryo

This award supports Dr. Edward Egelman of Yale University to collaborate in research with Dr. Jacques Dubochet of the Center for Electron Microscopy of the University of Lausanne, Switzerland. They are collaborating in a study of the RecA protein, the most intensively studied enzyme of general genetic recombination; their approach will also contribute substantially to improving techniques for the structural analysis of macromolecules in general. Dr. Egelman is a biophysicist who studies disordered and flexible macromolecules, specifically the RecA protein and actin. Traditional methods of structure determination, such as protein crystallography, have been generally unsuccessful with these molecules. Dr. Egelman has been examining them using computer-based image analysis of electron micrographs; he is developing a special facility for this purpose with support from the Instrumentation and Instrument Development Program of NSF. His particular expertise lies in graphics and software development; he depends on other laboratories for the electron micrographs to be studied. Dr. Dubochet has pioneered methods of cryo-electron microscopy of fully hydrated biological specimens. Only a handful of laboratories in the world are working in that field, and Dr. Dubochet is considered the leading expert on its use in imaging macromolecules. He will prepare frozen hydrated electron micrographs of RecA and other dynamical helical systems, which will provide one of the best means for both developing and demonstrating the effectiveness of Dr. Egelman's image processing hardware and software. Their complementary expertise and facilities will accelerate their mutual research on RecA and other dynamical protein polymers, and both laboratories will benefit from a reciprocal transfer of specialized techniques. The electron microscopy of frozen, fully hydrated biological specimens has emerged as one of the most promising new developments in structural biology in the past several years. It offers one of the most powerful methods available for visualizing disordered and dynamic helical protein polymers. It produces images of specimens with little or no damage or distortion from dehydration, heavy metal staining or electron dose. Dr. Egelman is developing instruments and programs to allow greatly accelerated analysis of electron micrographs, in particular to extract information about polymer dynamics and function from static images. His techniques for computer-aided enhancement and reconstruc- tion are even more necessary for frozen, hydrated specimens than for conventional electron micrographs, due to the problems of weak contrast and artifacts introduced by the optics of the electron microscope.

该奖项支持耶鲁大学的 Edward Egelman 博士与瑞士洛桑大学电子显微镜中心的 Jacques Dubochet 博士合作研究。 他们正在合作研究 RecA 蛋白，这是研究最深入的一般基因重组酶；他们的方法还将大大有助于改进一般大分子结构分析技术。 Egelman 博士是一位生物物理学家，研究无序且灵活的大分子，特别是 RecA 蛋白和肌动蛋白。 传统的结构测定方法，例如蛋白质晶体学，对于这些分子通常不成功。 埃格曼博士一直在使用基于计算机的电子显微照片图像分析来检查它们。在 NSF 仪器和仪器开发计划的支持下，他正在为此目的开发一种特殊设施。 他的特殊专长在于图形和软件开发；他依靠其他实验室来研究电子显微照片。 Dubochet 博士开创了完全水合生物样本的冷冻电子显微镜方法。 世界上只有少数实验室从事该领域的工作，而 Dubochet 博士被认为是该领域在大分子成像方面的领先专家。 他将准备 RecA 和其他动态螺旋系统的冷冻水合电子显微照片，这将为开发和展示 Egelman 博士的图像处理硬件和软件的有效性提供最佳手段之一。 他们互补的专业知识和设施将加速他们对 RecA 和其他动态蛋白质聚合物的共同研究，两个实验室都将受益于专业技术的相互转让。 冷冻、完全水合的生物标本的电子显微镜已成为过去几年结构生物学中最有前途的新发展之一。 它提供了可用于可视化无序和动态螺旋蛋白质聚合物的最强大方法之一。 它生成的样本图像很少或没有因脱水、重金属染色或电子剂量而造成的损坏或变形。 Egelman 博士正在开发仪器和程序，以极大地加速电子显微照片的分析，特别是从静态图像中提取有关聚合物动力学和功能的信息。 由于电子显微镜光学系统带来的对比度弱和伪影的问题，他的计算机辅助增强和重建技术对于冷冻、水合的标本比传统的电子显微照片更加必要。