Fast fragmentation method for characterization of peptides with labile PTMs

描述(申请人提供)：串联质谱仪(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影响。目前，基于ETD的MS分析作为一种独特的针对PTM的分析工具的适用性受到了限制，因为它的速度很慢，并且ETD无法有效地解离低电荷密度的蛋白质或肽。我们计划建造一种新型的ETD离子源，产生一束高能的负离子，以显著提高PTM检测蛋白质和多肽的效率和速度。为了展示基于ETD的高速率MS/MS分析，我们计划利用自行建造的配备多电极检测系统的台式FTMS仪器的能力，以高质量分辨率在短时间内记录快速ETD过程获得的MS/MS质谱图。 公共卫生相关性：在基于质谱学的复杂蛋白质混合物分析中，利用互补裂解方法是一种广泛使用的方法，可提供对蛋白质翻译后修饰的可靠识别。我们计划开发一种新的来源，提供快速碎片，这将大大改善和加速串联质谱仪分析的蛋白质翻译后修饰的表征。该源可以很容易地集成到各种串联质谱计的设计中。这将使从生物液或组织中更好地检测疾病特异性生物标记物。

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.