Midwest Consortium for High Resolution Cryoelectron Microscopy

中西部高分辨率冷冻电子显微镜联盟

基本信息

批准号：
10205086
负责人：
ESTHER BULLITT
金额：
$ 53.85万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10205086
关键词：
3-Dimensional Adhesions Alabama Antiviral Agents Back Bacterial Proteins Biological Process Boston Cell physiology Chemoreceptors Collaborations Collection Colorado Complex Cryoelectron Microscopy Data Data Collection Detection Development Disease Early Diagnosis Electron Beam Electron Microscope Electrons Engineering Ensure Equipment Faculty Fostering Freezing Funding Home Human Huntington Disease Image Institution Interruption Kansas Laboratories Light Macromolecular Complexes Manuals Medicine Methods Michigan Microscope Midwestern United States Missouri Motion Movement Performance Phase Productivity Proteins Recovery Research Research Personnel Resolution Resources Sampling Schedule Services Shipping Signal Transduction Site Structure Synaptic Vesicles Techniques Titan Training United States National Institutes of Health Universities Vermont Virginia Virus Virus Diseases Visit college contrast imaging cost data exchange detector experience faculty support flexibility high resolution imaging image processing improved insight instrument inter-institutional medical schools method development movie operation particle pathogenic bacteria portability protein complex public health relevance quantum reconstruction repaired supercomputer symposium three dimensional structure tomography tool transmission process

项目摘要

PROJECT SUMMARY Recently, single particle cryo-electron microscopy (cryo-EM) combined with 3-D reconstruction has emerged as a revolutionary tool for solving high-resolution 3-D structures of viruses and macromolecular complexes. The rapidly increasing number of near-atomic resolution (3-4Ǻ) structures solved using cryo-EM has allowed unprecedented atomic level understanding of fundamental cellular processes and viral infections. To obtain near-atomic resolution cryo-EM structures, requires the collection of high-resolution image data using state-of- the-art imaging resources, including both a high-end transmission electron microscope and a direct electron detector. The direct electron detectors not only improve image resolution and contrast, but also record movies for subsequent computational correction of electron beam-induced, sample movements during exposure. The increased image contrast and resolution are essential for solving near-atomic resolution structures of small protein complexes. However, the high cost to purchase and maintain a state-of-the-art cryo-EM resource, with both a high-end electron microscope and a direct electron detector, precludes many cryo-EM investigators from having access to these new techniques. Here, the creation of a Midwest Consortium for High- Resolution Cryo-electron Microscopy is proposed to provide access to such high-resolution data collection capability for cryo-EM laboratories without access to such resources. This consortium will consist of 4 investigators from the host institution (Purdue Univ.) which will maintain the high-resolution data collection resource (Titan Krios 300kV FEG microscope with phase plate, energy filter, and direct electron detector) and will provide services to 11 investigators from 10 partnering institutions (Boston Univ. School of Medicine, Michigan State Univ., Penn State College of Medicine, Rutgers Univ., Univ. of Alabama at Birmingham, Univ. of Colorado Boulder, Univ. of Kansas, Univ. of Missouri, Univ. of Vermont, and Virginia Commonwealth Univ.). The collective experience of the Purdue facility staff, faculty and onsite service engineer, in high-resolution cryo-EM imaging, will ensure that the facility operates at peak performance with minimal service interruptions. The high-resolution data collection capabilities established at Purdue and accessible to the partnering labs, will allow these investigators, who are all, except for the 3 new investigators, NIH-funded, to overcome the resolution barrier and facilitate discovery within their own cryo-EM projects on a range of structures, such as, bacterial pathogen adhesion proteins, human viruses, Huntington's Disease proteins, synaptic vesicle proteins, chemoreceptor signaling complex, etc. The Consortium will provide comprehensive support to the cryo-EM projects, including shipping samples, preparing sample grids, collecting high-resolution single particle and tomography images, processing raw movies, and transferring data back to the partners' labs. The data collection services will range from full data collection where partners simply mail in the samples and then receive the image data, to hands-on operations of the cryo-EM by partners trained by the host facility's staff.

项目摘要最近，已经出现了单个粒子冷冻电子显微镜（Cryo-EM）与3-D重建结合了作为解决病毒和大分子复合物的高分辨率3-D结构的革命性工具。使用冷冻EM求解的近原子分辨率（3-4ǻ）结构的迅速增加已允许对基本细胞过程和病毒感染的前所未有的原子水平的理解。获得接近原子分辨率的冷冻EM结构需要使用最先进的高分辨率图像数据收集艺术成像资源，包括高端传输电子显微镜和直接电子探测器。直接电子探测器不仅可以改善图像分辨率和对比度，还可以录制电影对于随后对电子束诱导的样品运动的计算校正。这增加图像对比度和分辨率对于解决小的接近原子分辨率至关重要蛋白质复合物。但是，购买和维护最先进的冷冻EM资源的高昂成本，高端电子显微镜和直接电子检测器都排除了许多冷冻EM研究者从访问这些新技术。在这里，建立了一个高级中西部财团提出了分辨率的低分辨率数据收集的分辨率冷冻电子显微镜冷冻EM实验室的能力，无需获得此类资源。该财团将包括4个主机机构（普渡大学）的调查人员将维持高分辨率数据收集资源（带相板，能量滤波器和直接电子检测器的泰坦krios 300kV FEG显微镜）和将为来自10个合作机构的11位调查人员提供服务（波士顿大学医学院，密歇根州立大学，宾夕法尼亚州立医学院，罗特大学，大学。阿拉巴马州伯明翰的阿拉巴马州。科罗拉多博尔德大学。堪萨斯大学。密苏里州，大学。佛蒙特州和弗吉尼亚联邦大学。普渡设施工作人员，教职员工和现场服务工程师的集体经验高分辨率冷冻EM成像将确保该设施在最小的服务中断时以高峰性能运行。在普渡大学建立和合作实验室可访问的高分辨率数据收集功能将除了三名由NIH资助的新调查人员以外，这些都是所有的调查员解决方案的障碍，并促进自己在自己的低温EM项目中发现的一系列结构，例如细菌病原体粘附蛋白，人类病毒，亨廷顿氏病蛋白，突触小泡蛋白，化学感受器信号传导复合物等。财团将为低温EM提供全面的支持项目，包括运输样品，准备样品网格，收集高分辨率的单个粒子和断层扫描图像，处理原始电影，然后将数据转移回合作伙伴的实验室。数据收集服务的范围从完整的数据收集中接收图像数据，并由由主机设施员工培训的合作伙伴进行冷冻EM的动手操作。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Integrating on-grid immunogold labeling and cryo-electron tomography to reveal photosystem II structure and spatial distribution in thylakoid membranes.

DOI：
10.1016/j.jsb.2021.107746
发表时间：
2021-09
期刊：
Journal of structural biology
影响因子：
3
作者：
Jiang J;Cheong KY;Falkowski PG;Dai W
通讯作者：
Dai W

Cryo-EM structures and functional characterization of homo- and heteropolymers of human ferritin variants.

DOI：
10.1038/s41598-020-77717-4
发表时间：
2020-11-26
期刊：
Scientific reports
影响因子：
4.6
作者：
Irimia-Dominguez J;Sun C;Li K;Muhoberac BB;Hallinan GI;Garringer HJ;Ghetti B;Jiang W;Vidal R
通讯作者：
Vidal R

Architecture of a complete Bce-type antimicrobial peptide resistance module.