Fast fragmentation method for characterization of peptides with labile PTMs

用于表征具有不稳定 PTM 的肽的快速裂解方法

• 批准号: 8315467
• 负责人: EUGENE MOSKOVETS
• 金额: $ 41.36万
• 依托单位: MASSTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-10 至 2014-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8315467
• 关键词: Affinity; Anions; Biological; Biological Markers; Biological Process; Cations; Cells; Cesium; Charge; Data; Detection; Disease; Dissociation; Electrodes; Electron Beam; Electron Transport; Electrons; Electrospray Ionization; Evaluation; Fast Electron; Fourier transform ion cyclotron resonance; Goals; Government; Hand; Housing; Hydrogen; Ions; Left; Legal; Liquid Chromatography; Liquid substance; Literature; Maps; Mass Spectrum Analysis; Methods; Metric; Modification; Molecular; Output; Peptide Fragments; Peptides; Persons; Phase; Phosphorylated Peptide; Phosphorylation; Post-Translational Protein Processing; Process; Production; Proteins; Proteomics; Protons; Reaction; Resolution; Role; Sensitivity and Specificity; Small Business Innovation Research Grant; Source; Speed; System; Techniques; Testing; Time; Tissues; United States National Institutes of Health; analytical tool; base; density; design; flexibility; improved; insight; instrument; ion source; mass spectrometer; novel; novel strategies; phase 2 study; protein complex; reaction rate; tandem mass spectrometry

DESCRIPTION (provided by applicant): Significant increase in the efficiency and speed of electron transfer dissociation (ETD) in tandem mass spectrometry (MS) will have a profound impact on its applications to the entire analytical field of characterization of posttranslational modifications (PTM) in proteins. At present, the applicability of ETD-based MS analysis as a unique analytical tool specifically targeting PTM is limited by its low speed and by inability of ETD to efficiently dissociate proteins or peptides with low charge density. We plan to build a novel ETD ion source generating an energetic beam of negative ions to significantly improve efficiency and speed of PTM detection of both proteins and peptides. To demonstrate high rate of MS/MS analysis based on ETD, we plan to utilize capabilities of house-built desktop FTMS instrument equipped with multi-electrode detection system, and record MS/MS mass spectra obtained from fast ETD process with high mass resolution and in a short time. PUBLIC HEALTH RELEVANCE: Utilizing complementary fragmentation methods is a broadly used approach in mass-spectrometry based analysis of complex protein mixtures providing reliable identification of posttranslation modifications in proteins. We plan to develop a novel source providing fast fragmentation that will substantially improve and accelerate characterization of posttranslation modifications in proteins analyzed in tandem mass spectrometers. The source can be easily incorporated into a design of a variety of tandem mass spectrometers. This will allow much better detection of disease-specific biomarkers from biological fluids or tissues.

描述（由申请人提供）：串联质谱（MS）中电子转移解离（ETD）效率和速度的显著提高将对其在蛋白质翻译后修饰（PTM）表征的整个分析领域的应用产生深远的影响。目前，基于ETD的质谱分析作为一种专门针对PTM的独特分析工具的适用性受到其低速度和ETD无法有效解离低电荷密度的蛋白质或肽的限制。我们计划建立一种新的产生负离子高能束的ETD离子源，以显著提高蛋白质和肽的PTM检测效率和速度。为了证明基于ETD的高MS/MS分析率，我们计划利用配备多电极检测系统的自制台式FTMS仪器的能力，在短时间内以高质量分辨率记录快速ETD过程中获得的MS/MS质谱。