Department of Chemistry
Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites
Determination of O-glycosylation sites in glycopeptides was developed by using two model compounds designed from mucin2 tandem repeat motif and erythropoietin. β-Elimination/addition reaction using dimethylamine on glycosylated site through a Michael-type condensation produced efficient deglycosylation with appropriate chemical modification. The use of dimethylamine was efficient to release the O-linked glycan in a reaction time period of 2-6 h at 55 °C. Peptide sequencing was then performed using the liquid chromatography/quadrupole time-of-flight mass spectrometry and MS-MS experiments. Interpretation of fragmentation pathways of the β-elimination/addition products enabled straightforward recognition of glycosylation site. Compared to the fragmentation of corresponding native peptides, mass shift of -18 Da or +27 Da was clearly observed for the two kinds of β-elimination/addition products of the glycosylated threonine. Dimethylamine was found to provide higher efficiency of β-elimination/addition than methylamine and ammonia. © 2008 Elsevier B.V. All rights reserved.
β-Elimination/addition, Glycopeptide, LC/Q-TOF MS, O-glycosylation site
Source Publication Title
Zheng, Yufang, Zhihong Guo, and Zongwei Cai. "Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites." Talanta 78.2 (2009): 358-363.