Advancing 'omics discovery via trapped ion mobility spectrometry

为了真正了解细胞的功能，我们必须充分了解大分子蛋白质机器和小分子之间的微妙相互作用，这些小分子调节它们的功能，生物合成，甚至捕捉它们短暂的相互作用。在这些小分子中，有附着在蛋白质特定位置的复杂支链糖，与细胞表面发现的蛋白质特异相互作用的脂类，以及调节细胞动态平衡和信号通路的代谢物。对这些生物分子的分析对于理解化学如何调节动态的生物过程至关重要，因为即使化学键的微小变化也可以对细胞功能产生重大影响。然而，由于它们固有的结构复杂性，这仍然是分析生物科学的主要挑战，因此我们对它们的生物学功能知之甚少。另一个技术挑战是描述瞬时蛋白质相互作用的能力，这可以通过使用快速反应的交联剂来捕捉，这种交联剂物理上永久地连接相互作用的伙伴，并促进他们的表征。质谱仪(MS)是分析这些化合物的主要手段，仪器技术的发展对于提高我们研究结构-功能关系和阐明它们在细胞、组织和整个生物体中的作用的能力是至关重要的。西德尼·布伦纳曾说过一句名言：“科学的进步依赖于新技术、新发现和新想法，很可能就是这样的顺序”，而我们的建议正是基于这一原则。这种新的质谱仪非常适合于这些生物分子的分析，并提供了几个优势，而不是目前最先进的平台，不能准确地识别复杂样品中存在的所有结构。该仪器的主要优点之一是如何通过在仪器内分离它们的离子物种来准确表征同分异构体结构，即具有相同原子组成但以不同方向组装的分子。这种质谱仪有着良好的记录，被广泛用于研究蛋白质；在这里，我们的目标是利用这项技术来研究寡糖、脂类、代谢物和化学交联的蛋白质片段。新仪器将整合到我们位于牛津大学化学系的实验室，致力于蛋白质复合体的结构分析-新仪器具有潜在的变革性，将对我们和我们的合作者的研究产生立竿见影的影响。它还将为BBSRC资助的研究人员提供急需的资源，以便在英国没有得到广泛支持(甚至没有)的专业领域进行实验。由于生物发现取决于获得最先进的设备，我们相信我们的提案将支持BBSRC的任务和优先事项，包括变革性技术和前沿生物科学。这种多用户和跨学科的仪器将在复杂小分子的分析以及我们确定其在健康和疾病过程中的生理相关功能的能力方面产生重大变化。

To truly appreciate how cells function, we must fully understand the delicate interplay between large macromolecular protein machines and small molecules that modulate their function, biosynthesis or even capturing their short-lived interactions. Among these small molecules are complex branched sugars attached to specific sites on a protein, lipids that specifically interact with proteins that are found on the surface of cells and metabolites that regulate cell homeostasis and signalling pathways. The analysis of these biomolecules is crucial for understanding how chemistry regulates dynamic biological processes as even small changes in a chemical bond can have significant effects in cellular function. However, owing to their inherent structural complexity, this remains a principal challenge in analytical bioscience and consequently we know comparatively little of their biological functions. Another technical challenge is the ability to characterise transient protein interactions, which can be captured using fast reacting crosslinking chemicals that physically attach interacting partners permanently and facilitates their characterisation. Mass spectrometry (MS) is the workhorse for the analysis of these compounds and technical developments in instrumentation are fundamental for progressing our ability to study structure-function relationships and elucidate their roles in cells, tissues and whole organisms. Sydney Brenner famously stated that "Progress in science depends on new techniques, new discoveries and new ideas, probably in that order" and our proposal is founded on this principle. This new mass spectrometer is perfectly suited for the analysis of these biomolecules and offers several advantages over the current state-of-the-art platforms that cannot accurately identify all structures present in a complex sample. One of the key benefits of the instrument is how isomeric structures, which are molecules with the same atomic composition but assembled in different orientations, can be accurately characterised by the separation of their ion species inside the instrument. This mass spectrometer has a proven track record and has been used extensively for studying proteins; here we aim to exploit this technology for the study of oligosaccharides, lipids, metabolites and chemically crosslinked protein fragments.The new instrument will be integrated into our laboratory in the Department of Chemistry in Oxford that is dedicated towards the structural analysis of protein complexes - the new instrument is potentially transformative and will have an immediate impact for our research and that of our collaborators. It will also provide a much-needed resource to BBSRC-funded researchers to undertake experiments in specialised fields which are not widely supported (or even absent) in the UK. As biological discoveries hinges on access to state-of-the-art equipment, we are confident our proposal will support BBSRC's remits and priorities, including transformative technologies and frontier bioscience. This multi-user and interdisciplinary instrument will ring a step-change in the analysis of complex small molecules and in our ability to determine their physiologically relevant functions in health and disease processes.