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

基本信息

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

项目摘要

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.

生命分子机器的功能与其在细胞内的环境密切相关。许多多蛋白复合物是动态的、脆弱的或与膜相关的，这使得使用传统的结构生物学方法（包括通过 X 射线晶体学、核磁共振或单粒子分析冷冻电镜进行蛋白质纯化和表征）进行有效研究具有挑战性或不可能。因此，原位研究分子机器对于我们充分了解其机制至关重要，并将为 21 世纪生物科学的发展奠定基础。此类基础工作还将应用于可视化疾病和病原体的原位机制、药物干预作用的细胞影响以及分子机器的合成设计（或现有的共同选择）。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，它利用电子冷冻显微镜的现有功能（例如，2017 年诺贝尔化学奖）对独特的生物样本进行成像，例如来自一系列角度来重建其 3D 结构并解析其分子成分：有效地进行“纳米级 CT 扫描”。这些 3D 图像或断层图解开了细胞的复杂性，否则在 2D 图像中细胞的复杂性会变得难以承受。随后可以对相似的结构（例如蛋白质复合物）进行排列、分类和平均，以获得其原位作用的 3D 结构快照。我们的建议是购买下一代“能量过滤器”，以实现常规电子冷冻断层扫描伦敦帝国学院。此次购买将通过一次升级购买推动帝国理工从“落后于时代”转变为“具有国际竞争力”，并促进我们各个 CoI 工作中已经萌芽的各种项目的发展。