Enabling ion Mobility Spectrometry/Mass Spectrometry Methods to Study Conformational Dynamics of Protein Systems

在化学系化学测量和成像项目的支持下，佛罗里达州立大学的Christian Bleiholder和他的研究小组正在开发新的方法来表征在生物细胞内发生的化学反应中起关键作用的蛋白质的结构动力学。这些新方法将结合并增强串联离子迁移谱法与质谱法结合计算化学方法的能力。因为这些方法可以提供与正常和异常细胞生物学相关的化学新知识，它们可能提供对细胞功能和细胞功能障碍的新见解，可能是提供洞察疾病进化的相关新工具。将通过与学术和工业合作者的相互作用以及通过网站和讲习班向潜在用户广泛传播思想和成果（包括软件工具）来加强影响。参与该项目的学生将接受仪器、计算方法、生物化学和复杂数据分析方面的培训。蛋白质的动态特性对蛋白质的化学反应有很大的影响。然而，在复杂样品或异质蛋白质系统的背景下研究蛋白质运动是具有挑战性的。离子迁移率谱法结合质谱法（IMS/MS）非常适合处理由于蛋白质形态的存在而引起的复杂性，并能够对蛋白质组进行系统的测量，但存在两个主要缺点：(1)在没有溶剂的情况下，溶液中蛋白质运动的哪些方面仍然不清楚；(2)从蛋白质的取向平均碰撞截面中明确地解释IMS/MS光谱是一项挑战。Bleiholder博士和他的团队将描述在没有溶剂的情况下，配体和非配体蛋白质系统的液相运动的哪些方面是保留的。这一目标是通过实验串联捕获IMS/MS （tandem-TIMS/MS）和Bleiholder实验室最近开发的计算结构松弛近似（SRA）方法来实现的。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 and Imaging Program in the Division of Chemistry, Christian Bleiholder and his research group at Florida State University are developing new approaches to characterize the structural dynamics of proteins that play critical roles in chemical reactions occurring within biological cells. These new approaches will combine and enhance the capabilities of tandem-ion mobility spectrometry coupled with mass spectrometry in conjunction with computational chemistry methods. Because these approaches can provide new knowledge on the chemistry associated with both normal and abnormal cell biology, they may offer novel insight into the functioning of cells and cell dysfunction, potentially a relevant new tool to provide insight into the evolution of disease. Impact will be enhanced through interactions with academic and industrial collaborators, and by broad dissemination of ideas and results (including software tools) to potential users through websites and workshops. Students working on the project will be trained in instrumentation, computational methods, biochemistry, and analysis of complex data. The dynamic nature of proteins contributes greatly to the chemical reactions of proteins. However, protein motions are challenging to study in the context of complex samples or heterogenous protein systems. Ion mobility spectrometry methods coupled with mass spectrometry (IMS/MS) are well-suited to handle the complexity arising from presence of proteoforms and enable systematic measurements of proteomes, but suffer from two major shortcomings: (1) It remains unclear which aspects of the solution phase protein motions are retained in the absence of solvent; and (2) it is challenging to unambiguously interpret IMS/MS spectra in terms of protein structures from their measured orientationally-averaged collision cross sections. Dr. Bleiholder and his team will characterize which aspects of solution phase motions of liganded and unliganded protein systems are retained in the absence of solvent. This objective is to be accomplished by means of the experimental tandem-trapped IMS/MS (tandem-TIMS/MS) and the computational Structure Relaxation Approximation (SRA) methods recently developed in the Bleiholder laboratory.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.