A next-generation energy filter for electron cryotomography at Imperial College

帝国理工学院用于电子冷冻断层扫描的下一代能量过滤器

• 批准号: BB/V019732/1
• 负责人: Morgan Beeby
• 金额: $ 53.04万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FV019732%2F1
• 关键词: next; generation; energy; filter; electron

The function of the molecular machines of life is intimately associated with their context inside cells. Many multiprotein complexes are dynamic, fragile, or membrane-associated, making them challenging or impossible to effectively study using the traditional structural biology approaches that involve protein purification and characterization by X-ray crystallography, nuclear magnetic resonance, or single particle analysis cryoEM. Studying molecular machines in situ is therefore crucial for us to fully understand their mechanisms and will form a foundation for the development of the biological sciences into the 21st century. Such foundational work will also have application in visualizing the mechanisms of diseases and pathogens in situ, the cellular ramifications of the action of pharmaceutical interventions, and the synthetic design (or co-option of existing) molecular machines. The only way to visualize the structural molecular biology of molecular machines in situ is by using electron cryotomography, which co-opts the existing power of electron cryomicroscopes (e.g., with the 2017 Nobel Prize in Chemistry) to image unique biological samples such as cells from a range of angles to reconstruct their 3-D structures and to resolve their molecular components: effectively a "nanoscale CT scan". These 3-D images, or tomograms, disentangle the complexity of the cell that would otherwise be overwhelming in a 2-D image; similar structures (e.g., protein complexes) can subsequently be aligned, classified, and averaged, to arrive at 3-D structural snapshots of their action in situ.Our proposal is to purchase a next generation "energy filter" to enable routine electron cryotomography at Imperial College London. This purchase will propel Imperial from being "behind the times" to "internationally competitive" with a single upgrade purchase and catalyse development of diverse projects already nascent in our various CoI's work.

生命的分子机器的功能与它们在细胞内的环境密切相关。许多多蛋白复合物是动态的，脆弱的，或膜相关的，使他们具有挑战性或不可能有效地研究使用传统的结构生物学方法，涉及蛋白质纯化和表征的X射线晶体学，核磁共振，或单粒子分析cryoEM。因此，原位研究分子机器对于我们充分理解它们的机制至关重要，并将为生物科学进入21世纪的发展奠定基础。这些基础工作还将应用于原位可视化疾病和病原体的机制、药物干预作用的细胞后果以及合成设计（或现有分子机器的协同选择）。原位可视化分子机器的结构分子生物学的唯一方法是使用电子冷冻断层扫描，其利用了电子冷冻显微镜的现有能力（例如，获得2017年诺贝尔化学奖）从一系列角度对独特的生物样品（如细胞）进行成像，以重建其三维结构并解析其分子组成：有效地进行“纳米级CT扫描”。这些3D图像或断层图像解开了细胞的复杂性，否则在2D图像中将是压倒性的;类似的结构（例如，蛋白质复合物）可以随后进行排列、分类和平均，以获得它们在原位作用的3-D结构快照。我们的建议是购买下一代“能量过滤器”，使伦敦帝国理工学院的常规电子冷冻断层扫描成为可能。此次收购将推动帝国从“落后于时代”到“国际竞争力”与一个单一的升级购买和催化发展的各种项目已经在我们的各种COI的工作初生。

项目成果

Evolution of a large periplasmic disk in Campylobacterota flagella facilitated efficient motility alongside autoagglutination

弯曲杆菌鞭毛中大型周质盘的进化促进了自凝集的有效运动

• DOI: 10.1101/2023.09.08.556628
• 发表时间: 2023
• 作者: Cohen E