Journal
ANALYTICAL CHEMISTRY
Volume 82, Issue 18, Pages 7588-7595Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac101306x
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Funding
- NSF [0945037]
- Div Of Industrial Innovation & Partnersh
- Directorate For Engineering [0945037] Funding Source: National Science Foundation
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This paper demonstrates the applications of a novel isobaric reagent, named deuterium (H-2) 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 H-2-associated isotope effects, such as partial loss of H-2 labels during tandem mass spectrometry (MS/MS) and H-2-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.
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