Combined Pulsed-Q Dissociation and Electron Transfer Dissociation for Identification and Quantification of iTRAQ-Labeled Phosphopeptides

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• 作者：Feng Yang ; Si Wu ; David L. Stenoien ; Rui Zhao ; Matthew E. Monroe ; Marina A. Gritsenko ; Samuel O. Purvine ; Ashoka D. Polpitiya ; Nikola Toli ; Qibin Zhang ; Angela D. Norbeck ; Daniel J. Orton ; Ronald J. M
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：May 15, 2009
• 年：2009
• 卷：81
• 期：10
• 页码：4137-4143
• 全文大小：186K
• 年卷期：v.81,no.10(May 15, 2009)
• ISSN：1520-6882

文摘

Here, we report a new approach that integrates pulsed Q dissociation (PQD) and electron transfer dissociation (ETD) techniques for confident and quantitative identification of iTRAQ-labeled phosphopeptides. The use of isobaric tags for relative and absolute quantification enables a high-throughput quantification of peptides via reporter ion signals in the low m/z range of tandem mass spectra. PQD, a form of ion trap collision activated dissociation, allows for detection of low mass-to-charge fragment ions, and electron transfer dissociation is especially useful for sequencing peptides that contain post-translational modifications. Analysis of the phosphoproteome of human fibroblast cells using a sensitive linear ion trap mass spectrometer demonstrated that this hybrid approach improves both identification and quantification of phosphopeptides. ETD improved phosphopeptide identification, while PQD provides improved quantification of iTRAQ-labeled phosphopeptides.