Deuterium isobaric amine-reactive tags for quantitative proteomics.
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.
Zhang
Junxiang
J
Institute of Bioscience and Biotechnology Research, University of Maryland, 9600 Gudelsky Drive, Rockville, Maryland 20850, USA.
Wang
Yan
Y
Li
Shuwei
S
D000588:Amines / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D003903:Deuterium / Q000737:chemistry*; D007202:Indicators and Reagents / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization
D000588:Amines; D007202:Indicators and Reagents; D010455:Peptides; D003903:Deuterium