Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites

Yufang Zheng, Zhihong Guo, Zongwei CAI*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)358-363
Number of pages6
JournalTalanta
Volume78
Issue number2
DOIs
Publication statusPublished - 30 Apr 2009

Scopus Subject Areas

  • Analytical Chemistry

User-Defined Keywords

  • Glycopeptide
  • LC/Q-TOF MS
  • O-glycosylation site
  • β-Elimination/addition

Fingerprint

Dive into the research topics of 'Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites'. Together they form a unique fingerprint.

Cite this