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: 10654823
• 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/10654823
• 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; biophysical techniques; care costs; drug development; drug discovery; improved; 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.

项目摘要/摘要 了解细胞过程和深度的途径，需要的方法不仅需要研究 各种蛋白质组件的组成和排列，但也阐明了它们的动态和 功能调节。实验策略提供了超出主要序列的结构信息； IE。， 高阶结构和蛋白质修饰，以产量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（即测量其本机溶液环境中的生物分子，以保存配体和其他 分子相互作用）和自上而下的MS/蛋白质组学将被提出并应用以促进 蛋白质和蛋白质组件的表征。耦合敏感电离，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，开斋节）和其他激活/解离技术超高分辨率质量 光谱法将提供一个实验平台，以实现完整的序列和蛋白质覆盖范围。这些 高级工具将被用来表征高分辨率的重要生物综合体 结构很难获得，包括G耦合蛋白受体（GPCR）和其他膜 蛋白质。探索翻译后修饰（PTM）与蛋白质成型之间的联系 神经退行性疾病（例如阿尔茨海默氏病（AD））的综合和毒性将通过 我们的高级MS方法。我们的实验策略广泛应用，并与许多 生物物理技术以实现大型和复杂的分子机器的详细结构研究，并 提供有关其动态的见解。本地自上而下的MS有望提高结构生物学并加速 药物发现与发育。改进基于MS的技术可以提高我们对 蛋白质和蛋白质机器如何驱动生物学。