Advancing Mass Spectrometry Analyses of Proteins, Assemblies, and Proteoforms

推进蛋白质、组装体和蛋白质形式的质谱分析

• 批准号: 10405346
• 负责人: Joseph A Loo
• 金额: $ 37.81万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10405346
• 关键词: Aging; Alzheimer' s Disease; Binding; Biological; Biological Process; Biology; Cell physiology; Complex; Coupling; Disease; Dissociation; Electrons; Environment; G-Protein-Coupled Receptors; Gases; Goals; Higher Order Chromatin Structure; Knowledge; Ligands; Link; Mass Spectrum Analysis; Measures; Membrane Proteins; Methodology; Methods; Modification; Molecular Machines; Neurodegenerative Disorders; Pathway interactions; Patient Care; Phase; Population; Post-Translational Protein Processing; Protein Analysis; Proteins; Proteomics; Regulation; Resolution; Site; Structure; Techniques; Technology; Toxic effect; base; biophysical techniques; care costs; drug development; drug discovery; insight; ionization; preservation; structural biology; tool; ultra high resolution

PROJECT SUMMARY/ABSTRACT Understanding cellular processes and pathways in depth requires methods that not only investigate the composition and arrangement of various protein assemblies, but that also elucidate their dynamics and functional regulation. Experimental strategies delivering structural information beyond primary sequence; i.e., higher-order structure and protein modifications, connect proteomics to structural biology in ways yielding potential insights into complex disease mechanisms. New techniques based on mass spectrometry (MS), native MS (i.e., measuring biomolecules in their native solution environment preserving ligand- and other molecular interactions), and top-down MS/proteomics will be advanced and applied to facilitate the characterization of proteins and protein assemblies. Coupling sensitive ionization, solution- and gas-phase separations, new electron-based dissociation methods (e.g., electron capture dissociation, ECD; electron ionization dissociation, EID) and other activation/dissociation techniques to ultra-high resolution mass spectrometry will provide an experimental platform for complete sequence and proteoform coverage. These advanced tools will be employed to characterize important biological complexes for which high-resolution structures have been difficult to obtain, including G-coupled protein receptors (GPCRs) and other membrane proteins. The exploration of links between post-translational modifications (PTMs) and proteoforms to the aggregation and toxicity of neurodegenerative diseases such as Alzheimer's disease (AD) will be furthered by our advanced MS methods. Our experimental strategies apply broadly and integrate with numerous biophysical techniques to enable the detailed structural study of large and complex molecular machines and to provide insights into their dynamics. Native top-down MS promises to advance structural biology and to hasten drug discovery and development. Improvements in MS-based technologies can advance our understanding of how proteins and protein machines drive biology.

项目概要/摘要 深入了解细胞过程和途径需要的方法不仅要研究 各种蛋白质组装体的组成和排列，但也阐明了它们的动态和 功能调节。提供一级序列之外的结构信息的实验策略； IE。， 高阶结构和蛋白质修饰，以产生的方式将蛋白质组学与结构生物学联系起来 对复杂疾病机制的潜在见解。基于质谱（MS）的新技术， 天然 MS（即在其天然溶液环境中测量生物分子，保留配体和其他 分子相互作用）和自上而下的 MS/蛋白质组学将得到推进和应用，以促进 蛋白质和蛋白质组装体的表征。耦合灵敏的电离、溶液和气相 分离、新的基于电子的解离方法（例如电子捕获解离、ECD；电子 电离解离 (EID) 和其他激活/解离技术以获得超高分辨率质量 光谱分析将为完整序列和蛋白质组覆盖提供一个实验平台。这些 将采用先进的工具来表征重要的生物复合物，这些生物复合物的高分辨率 结构很难获得，包括 G 偶联蛋白受体 (GPCR) 和其他膜 蛋白质。探索翻译后修饰（PTM）和蛋白质组之间的联系 阿尔茨海默病（AD）等神经退行性疾病的聚集和毒性将进一步加剧 我们先进的 MS 方法。我们的实验策略广泛适用并与众多 生物物理技术使大型复杂分子机器的详细结构研究成为可能 提供对其动态的见解。原生自上而下的 MS 有望推进结构生物学并加速 药物发现和开发。基于 MS 的技术的改进可以增进我们对 蛋白质和蛋白质机器如何驱动生物学。