Next-generation mass spectrometry of protein structure and interactions

蛋白质结构和相互作用的下一代质谱分析

• 批准号: EP/W021609/1
• 负责人: Justin Benesch
• 金额: $ 76.65万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FW021609%2F1
• 关键词: Next; generation; mass; spectrometry; protein

A major challenge at the interface of physical and life science is to understand how the function of proteins and other biomolecules is encoded in their structures and dynamics. These biomolecules are built from a small number of building blocks (such as amino acids and monosaccharides), which combine in a multitude of ways to create the vast diversity of interacting components that orchestrate processes necessary for life and responsible for malfunction in disease. A valuable tool for in-depth investigation of biological processes is mass spectrometry (MS), which provides fundamental information about the mass, identity and structure of biomolecules. With applications across almost all branches of science, mass spectrometers enable research into the mechanisms of health and disease, as well as drug discovery.To upgrade the capabilities of the advanced MS centre of excellence in the University of Oxford's Department of Chemistry, we seek to purchase a state-of-the-art mass spectrometer unrivalled in its ability to perform multidimensional analysis of protein stoichiometry, structure and dynamics. The instrument integrates MS with another high-resolution characterisation method, ion mobility spectrometry (IMS). IMS enables the measurement of molecular size based on the time taken to traverse a field of gas. In the cyclic implementation (cyclic IMS) we seek to obtain, ions can be passed indefinitely around a "race track", which dramatically increases the measurement resolution. The instrument will offer complementary capabilities (cyclic IMS, tandem MS, collision induced dissociation, electron capture dissociation and hydrogen deuterium exchange), which can be combined to tackle a variety of complex analytical problems. With cyclic IMS, for example, ions can be extracted from the ring, fragmented and then reinjected for further measurement. This capability makes it possible to probe the conformation-specific interactions of assemblies of macromolecules, revealing how subtle differences in structure correspond to differences in function.The first of its kind in the world, this instrument will bring a step change in our ability to interrogate complex biomolecular assemblies. It can perform all the measurements possible on our existing instruments, but at ten-fold higher resolution, and enable completely different types of experiments. This multi-user instrument will significantly boost the quality and capacity of research at the world-leading centre for gas-phase biophysics and structural biology. It will allow researchers within Oxford (across multiple departments and divisions) and the wider molecular characterisation community to capitalise on the latest developments in MS - to obtain measurements at unparalleled resolution, and probe the conformation-specific interactions of assemblies of proteins and other macromolecules at an unprecedented level of detail. These capabilities will make it possible to gain completely new insights into the mechanisms of function of macromolecules that play essential roles in health and disease, and often represent valuable therapeutic targets.

物理科学和生命科学的一个主要挑战是了解蛋白质和其他生物分子的功能如何在其结构和动力学中编码。这些生物分子是由少量的结构单元（如氨基酸和单糖）组成的，这些结构单元以多种方式联合收割机创造出各种各样的相互作用的成分，这些成分协调着生命所必需的过程，并导致疾病的功能障碍。深入研究生物过程的一个有价值的工具是质谱法（MS），它提供了有关生物分子的质量，身份和结构的基本信息。质谱仪应用于几乎所有的科学领域，使健康和疾病机制的研究以及药物发现成为可能。为了提升牛津大学化学系先进的MS卓越中心的能力，我们寻求购买一台最先进的质谱仪，其在对蛋白质化学计量，结构和动力学进行多维分析方面的能力无与伦比。该仪器将MS与另一种高分辨率表征方法离子迁移谱（IMS）相结合。IMS能够基于穿过气体场所花费的时间来测量分子大小。在我们寻求获得的循环实现（循环IMS）中，离子可以无限地绕着“赛道”通过，这大大提高了测量分辨率。该仪器将提供互补能力（循环IMS、串联MS、碰撞诱导解离、电子捕获解离和氢氘交换），这些能力可以结合起来解决各种复杂的分析问题。例如，使用循环IMS，离子可以从环中提取，破碎，然后重新注入以进行进一步测量。这一能力使得探测大分子组装体的构象特异性相互作用成为可能，揭示了结构上的细微差异如何对应于功能上的差异。这是世界上第一台此类仪器，它将为我们询问复杂生物分子组装体的能力带来一个飞跃。它可以在我们现有的仪器上进行所有可能的测量，但分辨率要高出十倍，并可以进行完全不同类型的实验。这一多用户仪器将显著提高世界领先的气相生物物理学和结构生物学中心的研究质量和能力。它将允许牛津大学（跨多个部门和部门）和更广泛的分子表征社区的研究人员利用MS的最新发展-以无与伦比的分辨率获得测量，并以前所未有的细节水平探测蛋白质和其他大分子组装的构象特异性相互作用。这些能力将使人们有可能获得对大分子功能机制的全新见解，这些大分子在健康和疾病中发挥着重要作用，并且通常代表着有价值的治疗靶点。