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

MRI：离子淌度表面诱导解离质谱仪的开发

• 批准号: 1342108
• 负责人: Vicki Wysocki
• 金额: $ 5.95万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1342108&HistoricalAwards=false
• 关键词: MRI; Development; Ion; Mobility; Surface

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%).

在国家科学基金重大研究仪器计划的支持下，一个项目的工作得以开始，该项目旨在设计和生产一种创新的离子迁移率表面诱导解离质谱仪，该仪器可以产生比目前商业仪器可能产生的关于大型非共价蛋白质复合体或其他大分子组件的更多结构信息。蛋白质通常通过在非共价多蛋白质复合体中共同工作来实现其功能。由于蛋白质复合体的总大小、生物基质的存在以及亚基之间非共价键的脆弱性质，表征蛋白质复合体中亚基的组成、数量、构象和空间排列是具有挑战性的。我们建议设计一种离子迁移率/表面诱导解离质谱仪，它可以测量大型非共价蛋白质络合物的m/z和形状(碰撞截面)，并有效地碎片形状选择的络合物来表征络合物的组成和结构。该开发项目将通过提供一种纳米电喷雾/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，该质谱仪可以在目前的仪器无法达到的水平上表征非共价蛋白质复合体。拟议工具的潜在用途范围很广：将要解决的生物学问题将侧重于确定非共价蛋白质和蛋白质-配基复合体中亚单位的化学计量和空间排列。开发项目本身将是将研究和教育结合起来的一个极好的工具，培养出学生和博士后助理，他们将学习如何在团队中工作，并将在生物大分子复合体的纯化、处理和分析以及仪器开发方面接受交叉培训。研究成果将在同行评议的文献、地方研讨会、工业合作者、国内和国际会议上、在加州大学国际分校化学和生物化学课程的研究生讲座上、以及由国际分会及其同事每年在美国质谱学学会年度全国会议上讲授的MS/MS短期课程中展示。国际学生联合会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%)。