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

在美国国家科学基金会重大研究仪器项目的支持下，该项目开始设计和生产一种创新的离子迁移率表面诱导解离质谱仪，该质谱仪可以产生比现有商用仪器更多的大型非共价蛋白质复合物或其他大分子组装的结构信息。蛋白质通常通过在非共价多蛋白复合物中协同工作来实现其功能。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，因为复合物的总大小、生物基质的存在以及亚基彼此之间非共价键的脆弱性。我们建议设计一种离子迁移率/表面诱导解离质谱仪，可以测量大型非共价蛋白复合物的m/z和形状（碰撞截面），并有效地片段形状选择复合物来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达到的水平上表征非共价蛋白质复合物。拟议仪器的潜在使用范围很广：将解决的生物学问题将集中在确定非共价蛋白质和蛋白质配体复合物中亚基的化学计量学和空间排列。该开发项目本身将是研究与教育相结合的绝佳工具，培养学生和博士后，他们将学习如何在团队中工作，并将在生物大分子复合物的纯化、处理和分析以及仪器开发方面进行交叉培训。研究成果将在同行评议的文献、本地研讨会、工业合作伙伴、国内和国际会议、PI在UA的化学和生物化学课程的研究生讲座，以及PI及其同事每年在美国质谱学会的年度全国会议上讲授的质谱/质谱短期课程中发表。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，这所学校的大多数学生是西班牙裔（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%).