Cryo EM/ET Core

冷冻 EM/ET 核心

• 批准号: 10506949
• 负责人: Peijun Zhang
• 金额: $ 33.88万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506949
• 关键词: Area; Attention; Capsid; Cells; Classification; Complement; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Electron Microscopy; Environment; Fluorescence Microscopy; HIV; HIV-1; Image; In Situ; Individual; Infection; Integration Host Factors; Ions; Label; Macromolecular Complexes; Methods; Microscopy; Nuclear Import; Nuclear Pore; Nuclear Pore Complex; Play; Process; Proteins; Resolution; Sampling; Scanning Electron Microscopy; Software Tools; Structure; Techniques; Technology; Viral; Virion; Work; X-Ray Crystallography; density; experience; flexibility; glycoprotein 41; inhibitor; light microscopy; nanometer resolution; new technology; nuclear pore complex protein 96; particle; protein complex; single molecule; small molecule inhibitor; software development; trafficking

Cryo-EM/ET Core Cryo-EM and cryo-ET are powerful techniques for high-resolution structural studies of proteins and protein complexes, complementing X-ray crystallography and NMR methods. Cryo-ET with subtomogram averaging and classification, in particular, has recently gained abundant attention as a prime method for characterization of macromolecular complexes that are intrinsically flexible and often challenging to purify for structure determination by single particle cryo-EM. Moreover, in situ cryo-ET can inform on structures of complexes within their native cellular environment, with near atomic resolution. The Cryo-EM/ET Core will play a key role in developing and applying these cutting-edge methods for structural analysis of heterogeneous HIV-1 viral components and their complexes with host cell factors and small molecule inhibitors. The Core will support PCHPI projects by providing structural information on assembly and maturation intermediates as well as capsid-host interactions during nuclear import and integration. Furthermore, the Core will offer new capabilities of cryo-EM and cryo-ET imaging of infectious samples in a biosafety level 3 (BSL-3) environment and develop new technologies and workflows for single-molecule correlation between cryo-ET and super-resolution fluorescence microscopy. These approaches will yield transformative structural and mechanistic information into the key steps of HIV-1 replication.

低温电子显微镜/等核心