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: 217080-2013
• 负责人: Konermann, Lars
• 金额: $ 7.21万
• 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: 2014
• 资助国家: 加拿大
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=546515
• 关键词: Protein; Structure; Dynamics; Studied; Mass

Most proteins in living organisms can perform their biological functions only after folding into a highly unique structure. Folding occurs spontaneously, driven by free energy gradients. Notably, the biologically active state remains quite dynamic, with structural fluctuations that are closely related to function. Misfolding can lead to Alzheimer's and many other diseases. Our group deciphers the mechanisms of protein folding and misfolding, as well as the interplay between structure, dynamics, and function. We have demonstrated international leadership in the development and application of novel protein mass spectrometry (MS) techniques. The current DG extends the reach and scope of these activities in a very significant way. (a) We will devise a method for elucidating the structures of the earliest folding intermediates. The combination of diffusive mixing with laser-induced oxidative labeling will improve the time resolution by three orders of magnitude compared to existing technologies. (b) We will employ oxidative labeling for mechanistic studies on a wide range of biologically important folding and self-assembly processes. (c) Hydrogen/deuterium exchange (HDX)/MS will be applied for gaining detailed insights into protein-ligand interactions. (d) Folding and dynamics of membrane proteins represent frontier areas that we will tackle using HDX/MS and oxidative labeling. (e) Shifting from traditional single-protein investigations to a "Systems Biology" perspective, we will be the first to apply structural/functional MS methods to protein networks, such as photosynthetic complexes. (f) Investigations on proteins in a solvent-free environment can reveal "intrinsic" biomolecular properties. Using a unique combination of gas-phase HDX, electron capture dissociation, ion mobility spectrometry, and computational modeling we will elucidate protein structures in the vacuum. Key to the success of all these activities is a carefully planned and executed strategy for HQP training. By becoming an integral part of our research enterprise HQP can develop an outstanding publication track record, as well as acquire the skills needed for successful careers in industry and academia.

活生物体中的大多数蛋白质只有在折叠成高度独特的结构后才能执行其生物学功能。折叠是由自由能梯度驱动的自发发生的。值得注意的是，生物学活性状态仍然是动态的，其结构波动与功能密切相关。错误折叠可能导致阿尔茨海默氏症和许多其他疾病。我们的小组解释了蛋白质折叠和错误折叠的机制，以及结构，动力学和功能之间的相互作用。我们已经证明了在新型蛋白质质谱法（MS）技术的开发和应用方面的国际领导。当前的DG以非常重要的方式扩展了这些活动的范围和范围。 （a）我们将设计一种阐明最早折叠中间体结构的方法。与现有技术相比，扩散混合与激光诱导的氧化标记的组合将提高三个数量级的时间分辨率。 （b）我们将使用氧化标记来进行机械研究，以实现广泛的生物学重要折叠和自组装过程。 （c）将应用氢/氘交换（HDX）/MS，以获得详细的洞察蛋白质 - 配体相互作用。 （d）膜蛋白的折叠和动力学代表我们将使用HDX/MS和氧化标记来应对的前沿区域。 （e）从传统的单蛋白研究转变为“系统生物学”的角度，我们将是第一个将结构/功能性MS方法应用于蛋白质网络的人，例如光合复合物。 （f）对无溶剂环境中蛋白质的研究可以揭示“固有”的生物分子特性。使用气相HDX，电子捕获解离，离子迁移率光谱法和计算建模的独特组合，我们将阐明真空中的蛋白质结构。所有这些活动成功的关键是对HQP培训的精心计划和执行的策略。通过成为我们研究企业HQP的不可或缺的一部分，可以发展出杰出的出版物记录，并获得行业和学术界成功职业所需的技能。