Next-generation mass spectrometry of protein structure and interactions

在物理和生命科学的交界处，一个主要的挑战是理解蛋白质和其他生物分子的功能是如何在它们的结构和动力学中编码的。这些生物分子是由少量的构件(如氨基酸和单糖)组成的，这些构件以多种方式结合在一起，创建了各种相互作用的成分，这些成分协调了生命所必需的过程，并对疾病的功能障碍负责。深入研究生物过程的一个有价值的工具是质谱仪(MS)，它提供关于生物分子的质量、身份和结构的基本信息。质谱计几乎应用于所有科学分支，使人们能够研究健康和疾病的机制，以及药物发现。为了提升牛津大学化学系高级MS卓越中心的能力，我们寻求购买一台最先进的质谱仪，其对蛋白质化学计量、结构和动力学的多维分析能力无与伦比。该仪器将MS与另一种高分辨率表征方法--离子迁移率光谱(IMS)相结合。IMS能够根据穿过气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一条赛道传递，这极大地提高了测量分辨率。该仪器将提供互补的功能(循环IMS、串联MS、碰撞诱导解离、电子俘获解离和氢氘交换)，它们可以结合在一起处理各种复杂的分析问题。例如，使用环状IMS，可以从环中提取离子，将其碎裂，然后重新注入以进行进一步测量。这种能力使探测大分子组装的特定构象的相互作用成为可能，揭示了结构上的细微差异与功能上的差异是如何对应的。这是世界上第一个此类仪器，它将使我们询问复杂生物分子组装的能力发生阶段性变化。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，并能够进行完全不同类型的实验。这一多用户仪器将大大提高世界领先的气相生物物理和结构生物学中心的研究质量和能力。它将允许牛津大学的研究人员(跨多个部门)和更广泛的分子表征社区利用MS的最新发展-以无与伦比的分辨率获得测量结果，并以前所未有的详细程度探索蛋白质和其他大分子组装的构象特定相互作用。这些能力将使人们有可能对大分子的功能机制获得全新的见解，这些大分子在健康和疾病中发挥着重要作用，通常代表着有价值的治疗靶点。

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.