Fast fragmentation method for characterization of peptides with labile PTMs

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影响。目前，基于ETD的MS分析作为特异性靶向PTM的独特分析工具的适用性受到其低速度和ETD不能有效解离具有低电荷密度的蛋白质或肽的限制。我们计划建立一种新型的ETD离子源，产生高能负离子束，以显着提高蛋白质和肽的PTM检测的效率和速度。为了证明基于ETD的MS/MS分析的高速率，我们计划利用配备多电极检测系统的自制台式FTMS仪器的功能，并在短时间内以高质量分辨率记录从快速ETD过程获得的MS/MS质谱。 公共卫生相关性：利用互补片段化方法是基于质谱分析的复杂蛋白质混合物中广泛使用的方法，其提供了蛋白质中翻译后修饰的可靠鉴定。我们计划开发一种新的来源，提供快速片段化，这将大大改善和加速表征的蛋白质翻译后修饰在串联质谱仪分析。该源可以很容易地结合到各种串联质谱仪的设计中。这将允许从生物流体或组织中更好地检测疾病特异性生物标志物。

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.