CAREER: Ion Mobility Spectrometry - Mass Spectrometry for de novo Protein Structure Elucidation

在化学系化学测量和成像计划的支持下，以及生命过程化学计划(化学系)和分子生物物理组(分子和细胞生物科学系)的共同资助下，佛罗里达州立大学的Christian Bleihold教授和他的团队正在开发新的工具来表征在正常和异常(疾病)细胞的化学中发挥关键作用的蛋白质的结构。这些工具结合并增强了离子迁移率和质谱学测量的能力，可能提供迄今为止用现有工具无法获得的新的生化见解。这反过来可能为细胞的功能和疾病的原因提供新的见解。通过与学术和工业合作者的互动，以及通过网站、研讨会和教程向潜在用户广泛传播想法和成果(包括软件工具)，提高了影响力。参与这些项目的学生接受了仪器、计算方法、生物化学和复杂数据分析方面的培训。蛋白质及其组装的瞬时填充构象在细胞的功能(和功能失调)中起着重要作用。近年来，离子迁移率光谱-质谱仪(IMS-MS)方法被证明能够研究这种瞬时构象，但仍然存在两个主要缺陷：IMS-MS阐明的蛋白质结构与溶液中生物活性结构之间的关系尚不清楚，以及详细的蛋白质结构很大程度上无法从IMS-MS数据中获得。布莱霍尔德博士和他的团队正在通过开发新的实验和计算方法来获取和解释时间分辨的IMS-MS数据，以应对这些挑战。为了评估从IMS-MS数据中阐明蛋白质结构的准确性，该团队正在评估天然蛋白质结构可以在气相中保留多长时间和程度。正在开发和评估计算方法，因为它们能够仅根据蛋白质的横截面分布来识别蛋白质的结构。最后，开发的计算方法以非专家易于使用的形式传播，以便使更广泛的IMS-MS社区能够准确地阐明蛋白质结构。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry and co-funding from the Chemistry of Life Processes Program (Division of Chemistry) and the Molecular Biophysics Cluster (Division of Molecular and Cellular Biosciences), Professor Christian Bleiholder and his group at Florida State University are developing new tools to characterize the structures of proteins that play critical roles in the chemistry of both normal and abnormal (diseased) cells. These tools - which combine and enhance the capabilities of ion mobility and mass spectrometric measurements - may provide new biochemical insights which have thus far not been possible to obtain with existing tools. This in turn may offer new insights into the functioning of cells and the causes of disease. Impact is enhanced through interactions with academic and industrial collaborators, and by broad dissemination of ideas and results (including software tools) to potential users through websites, workshops, and tutorials. Students working on the projects are trained in instrumentation, computational methods, biochemistry, and analysis of complex data. Transiently populated conformations of proteins and their assemblies play important roles in the function (and misfunction) of cells. Ion mobility spectrometry - mass spectrometry (IMS-MS) methods have recently been shown to be capable of studying such transient conformations but still suffer from two major shortcomings: the relationship between the protein structure elucidated by IMS-MS and the biologically active structure in solution is poorly understood, and detailed protein structures are largely inaccessible from IMS-MS data. Dr. Bleiholder and his team are approaching these challenges by developing new experimental and computational approaches for obtaining and interpreting time-resolved IMS-MS data. To assess the accuracy of protein structure elucidation from IMS-MS data, the team is assessing how long and to what extent native protein structures can be retained in the gas phase. Computational methods are being developed and assessed for their ability to identify the structure of a protein from only its cross section distribution. Finally, the developed computational methods is disseminated in a form that is easy to use by non-experts in order to enable a wider IMS-MS community to elucidate protein structures accurately.