Cryo EM/ET Core

冷冻EM/ET核心 Cryo-EM和Cryo-ET是研究蛋白质和蛋白质高分辨率结构的强有力技术 配合物，补充X射线晶体学和NMR方法。Cryo-ET，子断层图像平均， 分类，特别是，最近获得了大量的关注，作为表征的主要方法， 大分子复合物具有固有的柔性，并且通常难以纯化以进行结构测定 单粒子冷冻电镜此外，原位冷冻ET可以告知复合物的结构， 细胞环境，具有接近原子的分辨率。Cryo-EM/ET核心将在开发和 将这些尖端方法应用于异质性HIV-1病毒组分及其 与宿主细胞因子和小分子抑制剂的复合物。核心将通过提供以下服务来支持PCHPI项目： 组装和成熟中间体的结构信息，以及在组装过程中的captant-宿主相互作用， 核进口和一体化。此外，Core还将提供新的冷冻EM和冷冻ET成像功能 在生物安全3级（BSL-3）环境中检测感染性样本，并开发新技术和工作流程 冷冻ET和超分辨率荧光显微镜之间的单分子相关性。这些 这些方法将为HIV-1的关键步骤提供变革性的结构和机制信息， 复制的

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.