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
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Zheng, Y., Guo, Z., & Cai, Z. (2009). Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites. Talanta, 78 (2), 358-363. https://doi.org/10.1016/j.talanta.2008.11.026