Deuterium isobaric amine-reactive tags for quantitative proteomics.

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TitleDeuterium isobaric amine-reactive tags for quantitative proteomics.
Publication TypeJournal Article
Year of Publication2010
AuthorsZhang, J, Wang, Y, Li, S
JournalAnal Chem
Volume82
Issue18
Pagination7588-95
Date Published2010 Sep 15
ISSN1520-6882
KeywordsAmines, Animals, Cattle, Deuterium, Indicators and Reagents, Peptides, Phosphorylation, Proteomics, Spectrometry, Mass, Electrospray Ionization
Abstract

This paper demonstrates the applications of a novel isobaric reagent, named deuterium ((2)H) isobaric amine-reactive tag (DiART), for quantitative proteomics. Peptides labeled with DiART were analyzed using an electrospray ionization (ESI)-based LTQ-Orbitrap mass spectrometer. Our data showed that (2)H-associated isotope effects, such as partial loss of (2)H labels during tandem mass spectrometry (MS/MS) and (2)H-related chromatographic shift, were either not observed or negligible. With the use of a hybrid collision-induced dissociation (CID)-higher energy C-trap dissociation (HCD) acquisition method, we were able to identify DiART-labeled peptides with high confidence and quantify them with high accuracy. Furthermore, we adopted a hybrid electron-transfer dissociation (ETD)-HCD acquisition protocol and developed a novel data analysis approach to measure phosphorylation of peptides. Our results showed DiART had excellent performance on LTQ-Orbitrap instruments and provided a cost-effective technique for large-scale quantitative proteomics measurements.

DOI10.1021/ac101306x
Alternate JournalAnal. Chem.
PubMed ID20715779