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.

我们的实验室探讨了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，我们开发并应用了提供独特见解的新型电喷雾电离（ESI）质谱（MS）技术。我们通过光谱和尖端分子动力学（MD）模拟完成所得数据。过去的NSERC资金使我们的团队进入了全球领导地位。该DG将通过关注三个相互联系的主题1。蛋白质动力学和功能。氢/氘交换（HDX）/MS是探测溶液中蛋白质动力学的重要工具。柔性段会经历快速的HDX，而折叠区域的剥离却很慢。我们将对HDX保护效应有很大的改进，为基于HDX/MS的结构建模工作奠定了基础。 HDX/MS将发现关键蛋白（包括治疗抗体）的折叠机制，内部动力学和降解现象。关于基于蛋白质的分子机器的HDX/MS研究将阐明与润滑，摩擦，变形和功率传播有关的方面。这些结果将与纳米技术中的未来应用有关2。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当前现有的氧化物标记方法的局限性，从而将前所未有的见解延伸到蛋白质折叠中间体的结构中。我们还将阐明蛋白质氧化在细胞色素C的过氧化物酶激活中的作用，该主题与癌细胞的凋亡消除有关。3。 ESI机理和气相蛋白。 “天然” ESI-MS广泛用于探测蛋白质和生物复合物。但是，尚不清楚电喷雾分析物在气相中保留其结构的程度。我们将审查以下假设：广泛的盐桥网络将动力学稳定性赋予天然样气相蛋白。此外，我们的工作将揭示蛋白质是否可以在气相中折叠，从而阐明水在生物分子自组装中的作用。新颖的计算途径将产生ESI过程的“分子电影”，将天然ESI-MS领域放置在实心理论/计算基础上。HQP训练。该提案围绕精心计划和执行的HQP培训策略。成为我们团队的一员为HQP提供了出色的往绩，其中包括知名出版物，主要奖项和享有声望的科学。与其他研究小组和机构的合作极大地促进了刺激的跨学科环境。这种培训背景为行业或学术界繁荣的职业提供了跳板。