Protein Structure and Dynamics Studied by Mass Spectrometry

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发挥其生物学功能。折叠在自由能梯度的驱动下自发发生。值得注意的是，生物活性状态仍然是动态的，结构波动与功能密切相关。错误折叠会导致阿尔茨海默氏症和许多其他疾病。我们的团队破译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。我们在新型蛋白质质谱（MS）技术的开发和应用方面表现出国际领先地位。现任总干事以非常重要的方式扩大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氧化标记的结合将使时间分辨率提高三个数量级。(b)我们将在广泛的生物学上重要的折叠和自组装过程的机制研究中使用氧化标记。(c)氢/氘交换(HDX)/质谱将用于获得蛋白质-配体相互作用的详细见解。(d)膜蛋白的折叠和动力学是我们将使用HDX/MS和氧化标记来解决的前沿领域。(e)从传统的单蛋白研究转向“系统生物学”的观点，我们将率先将结构/功能质谱方法应用于蛋白质网络，如光合复合体。(f)在无溶剂环境中对蛋白质进行研究可以揭示“内在的”生物分子特性。利用气相HDX，电子捕获解离，离子迁移率光谱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精心策划和执行HQP培训战略。通过成为我们研究事业的一个组成部分，HQP可以发展出杰出的出版记录，并获得在工业界和学术界成功职业所需的技能。

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.