|Title||Deuterium isobaric amine-reactive tags for quantitative proteomics.|
|Publication Type||Journal Article|
|Year of Publication||2010|
|Authors||Zhang, J, Wang, Y, Li, S|
|Date Published||2010 Sep 15|
|Keywords||Amines, Animals, Cattle, Deuterium, Indicators and Reagents, Peptides, Phosphorylation, Proteomics, Spectrometry, Mass, Electrospray Ionization|
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.
|Alternate Journal||Anal. Chem.|