Bridging the Gap Between Observables from Ion Mobility Mass Spectrometry and the Structures of Native Proteins

在化学系化学测量与成像（CMI）项目的支持和分子与细胞生物科学系的部分共同资助下，布什教授和他在华盛顿大学的团队正在开发表征生物分子结构的新方法。布什实验室正在开发的最先进的技术为这些测量提供了新的方法，解决了对生物分子的结构、组装、异质性、质量和相似性的快速表征的未满足需求。新的理论和统计方法也正在开发和应用，以深入了解所研究的蛋白质的气相测量和溶液相结构之间的关系。这些知识将有助于回答有关蛋白质和蛋白质复合物的结构和功能的重要问题，这对于理解生命和疾病的分子基础非常重要。在布什实验室工作的学生接受测量科学、数据科学和通信方面的培训，为他们在各种各样的职业生涯中取得成功做好准备，从而提高美国在分析仪器和生物技术领域的经济竞争力。该小组还在开发教学模块，使广大学生能够接触到他们研究的基本概念。尽管离子迁移率（IM）质谱（MS）具有许多特性，使其能够很好地满足结构生物学和生物物理学中尚未满足的需求，但对溶液和气相结构保真度的担忧抑制了IM-MS技术的广泛采用，并降低了基于IM-MS的结构模型的信心。布什小组正致力于(1)表征电荷状态对蛋白质离子结构的影响；(2)表征蛋白质离子重排动力学；(3)利用现代统计方法增加当前IM实验和计算的信息量。本研究的结果将通过提供对原生类离子的结构和动力学的详细了解，开发更准确的方法将IM-MS实验中的可观测值转化为结构建模的约束，并开发一种将结构模型转化为可观测值的准确预测的通用方法，解决IM-MS社区的迫切需求。这反过来将使IM-MS在混合结构生物学项目和倡议中发挥更大的作用。将开发虚拟实验室，使学生能够接触到IM-MS技术，即使在没有IM-MS仪器的地方。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Measurement & Imaging (CMI) Program in the Division of Chemistry and partial co-funding from the Division of Molecular and Cellular Biosciences, Professor Bush and his group at the University of Washington are developing new methods to characterize the structures of biological molecules. State-of-the-art techniques being developed in the Bush lab enable new approaches to these measurements, addressing unmet needs for rapid characterization of the structures, assembly, heterogeneity, quality, and similarity of biological molecules. New theoretical and statistical methods are also being developed and applied to gain insight into the relationship between the gas-phase measurements and the solution-phase structures of the proteins studied. This knowledge will help answer important questions about the structure and function of proteins and protein complexes, which is important for understanding the molecular basis of life and disease. Students working in the Bush lab are trained in measurement science, data science, and communication, preparing them for success in a wide variety of careers, thereby increasing the economic competitiveness of the United States in the fields of analytical instrumentation and biotechnology. The group is also developing teaching modules to enable exposure of a wide range of students to the concepts underlying their research.Although ion mobility (IM) mass spectrometry (MS) has many attributes that make it well positioned to fulfill unmet needs in structural biology and biophysics, concerns about the fidelity of solution- and gas-phase structures have inhibited the broader adoption of IM-MS technologies and reduced the confidence in structural models based on IM-MS. The Bush group is working to (1) characterize the effects of charge state on the structures of protein ions; (2) characterize the dynamics of protein ions rearrangements; and (3) increase the information content of current IM experiments and calculations using modern statistical methods. The outcomes of this research will address urgent needs in the IM-MS community by providing detailed knowledge of the structures and dynamics of native-like ions, developing more accurate methods for translating observables from IM-MS experiments into restraints for structural modeling, and developing a general method for translating structural models into accurate predictions of observables. This in turn will enable IM-MS to play a greater role in hybrid structural biology projects and initiatives. Virtual labs will be developed to enable exposure of students to IM-MS techniques even where IM-MS instrumentation is not available.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.