MRI: Development of an Ion Mobility Surface-Induced Dissociation Mass Spectrometer

国家科学基金会主要研究仪器计划的支持，使工作开始的一个项目，设计和生产一个创新的离子迁移率表面诱导解离质谱仪，可以产生更多的结构信息大非共价蛋白质复合物或其他大分子组装比目前的商业仪器。蛋白质通常通过在非共价多蛋白质复合物中一起工作来实现其功能。表征蛋白质复合物中亚基的组成、数量、构象和空间排列是具有挑战性的，因为总复合物的大小、生物基质的存在以及亚基彼此非共价键合的脆弱性质。我们建议设计一个离子迁移率/表面诱导解离质谱仪，可以测量的m/z和形状（碰撞截面）的大型非共价蛋白质复合物和有效的片段形状选择的复合物，以表征复合物的组成和结构。该开发项目将通过提供纳米电喷雾/离子迁移率/表面诱导解离串联质谱仪来改善结构生物学研究的基础设施，该质谱仪可以在当前仪器无法实现的水平上表征非共价蛋白质复合物。拟议的仪器的潜在用途的范围是广泛的：生物学问题，将被解决将集中在非共价蛋白质-蛋白质和蛋白质-配体复合物中的亚单位的化学计量和空间排列的确定。 开发项目本身将是一个很好的工具，结合研究和教育，生产学生和博士后助理谁将学习如何在团队中工作，谁将在生物大分子复合物的纯化，处理和分析以及仪器开发的交叉培训。研究结果将在同行评议的文献中，在当地研讨会上，向工业合作者，在国家和国际会议上，由PI在UA的化学和生物化学课程的研究生讲座中，以及由PI和同事每年在美国质谱学会年度全国会议上教授的MS/MS短期课程中介绍。PI的研究小组还将与图森市曼斯菲尔德中学的两名当地六年级科学教师合作，这所学校的大部分学生是西班牙裔（68%）。

Support from the National Science Foundation Major Research Instrumentation Program has allowed work to begin on a project to design and produce an innovative ion mobility surface-induced dissociation mass spectrometer that can produce much more structural information on large non-covalent protein complexes or other macromolecular assemblies than is possible with present commercial instrumentation. Proteins often carry out their functions by working together in non-covalent multi-protein complexes. Characterization of the composition, numbers, conformations, and spatial arrangements of the subunits in a protein complex is challenging because of the total complex size, the presence of a biological matrix, and the fragile nature of the non-covalent bonding of the subunits to each other. We propose to design an ion mobility/surface-induced dissociation mass spectrometer that can measure the m/z and shapes (collision cross sections) of large non-covalent protein complexes and effectively fragment shape-selected complexes to characterize the composition and structure of the complex. The development project will improve the infrastructure for structural biology research by providing a nanoelectrospray/ion mobility/surface-induced dissociation tandem mass spectrometer that can characterize non-covalent protein complexes at a level not possible with current instrumentation. The scope of potential use of the proposed instrument is broad: biological problems that will be addressed will focus on determination of the stoichiometry and spatial arrangement of sub-units in non-covalent proteinprotein and protein-ligand complexes. The development project itself will be an excellent vehicle for combining research and education, producing students and postdoctoral associates who will learn how to work in teams and who will be cross-trained in the purification, handling, and analysis of biological macromolecular complexes and in instrument development. The research results will be presented in the peer-reviewed literature, at local seminars, to industrial collaborators, at national and international conferences, in graduate lectures by the PI at UA in Chemistry and Biochemistry courses, and in the MS/MS short course taught by the PI and colleagues each year at the annual national meeting of the American Society for Mass Spectrometry. The PI's research group will also work with two local 6th grade science teachers at Mansfeld Middle School in Tucson - a school where the majority of students are Hispanic (68%).