Protein Structure and Dynamics Studied by Mass Spectrometry

通过质谱研究蛋白质结构和动力学

• 批准号: RGPIN-2018-04243
• 负责人: Konermann, Lars
• 金额: $ 8.81万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=747951
• 关键词: Protein; Structure; Dynamics; Studied; Mass

Our laboratory explores the mechanisms of protein folding and misfolding, as well as the relationship between protein dynamics, interactions, and function. For this purpose, we develop and apply novel electrospray ionization (ESI) mass spectrometry (MS) techniques that provide unique insights. We complement the resulting data with optical spectroscopy and cutting-edge molecular dynamics (MD) simulations. Past NSERC funding has allowed our group to move into a global leadership position. This DG will further extend the reach and scope of our activities by focusing on three interconnected themes.1. Protein Dynamics and Function. Hydrogen/deuterium exchange (HDX)/MS is an important tool for probing protein dynamics in solution. Flexible segments undergo rapid HDX, while deuteration in folded regions is slow. We will develop a greatly improved understanding of HDX protection effects, providing the foundation for HDX/MS-based structural modeling efforts. HDX/MS will uncover the folding mechanisms, internal dynamics, and degradation phenomena of key proteins, including therapeutic antibodies. HDX/MS studies on protein-based molecular machines will elucidate aspects related to lubrication, friction, deformation, and power transmission. These results will be relevant for future applications in nanotechnology.2. Analytical and Biomedical Aspects of Protein Oxidation. Innovative radical chemistry will overcome limitations of currently existing oxidative labeling methods, paving the way towards unprecedented insights into the structures of protein folding intermediates. We will also elucidate the role of protein oxidation for the peroxidase activation of cytochrome c, a topic that is related to the apoptotic elimination of cancer cells.3. ESI Mechanisms and Gas Phase Proteins. “Native” ESI-MS is widely used for probing proteins and biological complexes. However, it remains unclear to what extent electrosprayed analytes retain their structures in the gas phase. We will scrutinize the hypothesis that extensive salt bridge networks impart kinetic stability to native-like gas phase proteins. In addition, our work will reveal whether proteins can fold in the gas phase, shedding light on the role of water during biomolecular self-assembly. Novel computational avenues will yield “molecular movies” of the ESI process, placing the field of native ESI-MS on solid theoretical/computational footing.HQP Training. This proposal revolves around a carefully planned and executed HQP training strategy. Being part of our team provides HQP with an exceptional track record that includes high profile publications, major awards, and prestigious scholarships. Collaborations with other research groups and institutions greatly contribute to a stimulating interdisciplinary environment. This training background provides a springboard for prosperous careers in industry or academia.

我们的实验室探索蛋白质折叠和错误折叠的机制，以及蛋白质动力学，相互作用和功能之间的关系。为此，我们开发和应用新的电喷雾电离（ESI）质谱（MS）技术，提供独特的见解。我们用光谱学和尖端的分子动力学（MD）模拟来补充所得到的数据。过去的NSERC资助使我们的团队进入了全球领导地位。该总干事将进一步扩大我们活动的范围和广度，重点关注三个相互关联的主题。蛋白质动力学与功能氢/氘交换（HDX）/MS是探测溶液中蛋白质动力学的重要工具。柔性片段经历快速HDX，而折叠区域中的氘化是缓慢的。我们将大大提高对HDX保护效果的理解，为基于HDX/MS的结构建模工作提供基础。HDX/MS将揭示关键蛋白质（包括治疗性抗体）的折叠机制、内部动力学和降解现象。 HDX/MS对蛋白质分子机器的研究将阐明与润滑、摩擦、变形和动力传输相关的方面。这些结果将与未来在纳米技术中的应用有关。蛋白质氧化的分析和生物医学方面。创新的自由基化学将克服现有氧化标记方法的局限性，为前所未有的蛋白质折叠中间体结构的深入研究铺平道路。我们还将阐明蛋白质氧化对细胞色素c的过氧化物酶激活的作用，这是一个与癌细胞凋亡消除相关的主题。ESI机制和气相蛋白质。“天然”ESI-MS广泛用于探测蛋白质和生物复合物。然而，目前尚不清楚电喷雾分析物在气相中保留其结构的程度。我们将仔细研究的假设，广泛的盐桥网络赋予动力学稳定性天然样气相蛋白质。此外，我们的工作将揭示蛋白质是否可以在气相中折叠，从而揭示水在生物分子自组装过程中的作用。新的计算途径将产生ESI过程的“分子电影”，将天然ESI-MS领域置于坚实的理论/计算基础上。HQP培训。该建议围绕着一个精心计划和执行的HQP培训策略。作为我们团队的一员，HQP拥有卓越的业绩记录，包括备受瞩目的出版物，主要奖项和着名奖学金。与其他研究团体和机构的合作极大地促进了激励跨学科的环境。这种培训背景为工业界或学术界的繁荣事业提供了跳板。