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，我们开发并应用了新型电喷雾电离 (ESI) 质谱 (MS) 技术，提供了独特的见解。我们利用光谱学和尖端分子动力学 (MD) 模拟来补充所得数据，使我们的团队能够跻身全球领导地位，重点关注三个领域。 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。了解 HDX 保护作用，为基于 HDX/MS 的结构建模工作奠定基础，HDX/MS 将揭示关键蛋白质的折叠机制、内部动力学和降解抗体现象，包括基于蛋白质的治疗研究。分子机器将阐明与润滑、摩擦、变形和动力传输相关的方面。这些结果将与纳米技术的未来应用相关。2. 创新的自由基化学将克服现有氧化标记方法的局限性。 ，为深入了解蛋白质折叠中间体的结构铺平了道路，我们还将阐明蛋白质氧化对于细胞色素 c 过氧化物酶激活的作用。与癌细胞凋亡消除相关的主题。3.“天然”ESI-MS 广泛用于探测蛋白质和生物复合物，但目前尚不清楚电喷雾分析物在多大程度上保留了其结构。我们将仔细研究广泛的盐桥网络赋予类天然气相蛋白质动力学稳定性的假设，此外，我们的工作还将揭示蛋白质是否可以在气相中折叠，从而揭示这一点。关于水在生物分子自组装过程中的作用的新计算途径将产生 ESI 过程的“分子电影”，为本地 ESI-MS 领域奠定坚实的理论/计算基础。该提案围绕精心计划的内容。并执行 HQP 培训战略，为 HQP 提供了卓越的业绩记录，其中包括高知名度的出版物、重大奖项以及与其他研究小组和机构的合作，这为激励性的研究做出了贡献。这种培训背景为工业界或学术界的繁荣职业生涯提供了跳板。