Analytical and Bioanalytical Chemistry

, Volume 409, Issue 2, pp 579–588 | Cite as

Isotope-targeted glycoproteomics (IsoTaG) analysis of sialylated N- and O-glycopeptides on an Orbitrap Fusion Tribrid using azido and alkynyl sugars

  • Christina M. Woo
  • Alejandra Felix
  • Lichao Zhang
  • Joshua E. Elias
  • Carolyn R. Bertozzi
Research Paper
Part of the following topical collections:
  1. Glycomics, Glycoproteomics and Allied Topics

Abstract

Protein glycosylation is a post-translational modification (PTM) responsible for many aspects of proteomic diversity and biological regulation. Assignment of intact glycan structures to specific protein attachment sites is a critical step towards elucidating the function encoded in the glycome. Previously, we developed isotope-targeted glycoproteomics (IsoTaG) as a mass-independent mass spectrometry method to characterize azide-labeled intact glycopeptides from complex proteomes. Here, we extend the IsoTaG approach with the use of alkynyl sugars as metabolic labels and employ new probes in analysis of the sialylated glycoproteome from PC-3 cells. Using an Orbitrap Fusion Tribrid mass spectrometer, we identified 699 intact glycopeptides from 192 glycoproteins. These intact glycopeptides represent a total of eight sialylated glycan structures across 126 N- and 576 O-glycopeptides. IsoTaG is therefore an effective platform for identification of intact glycopeptides labeled by alkynyl or azido sugars and will facilitate further studies of the glycoproteome.

Keywords

Glycoproteomics Chemical proteomics LC-MS/MS Metabolic labeling Sialic acid 

Supplementary material

216_2016_9934_MOESM1_ESM.pdf (3.3 mb)
ESM 1(PDF 3393 kb)
216_2016_9934_MOESM2_ESM.xlsx (571 kb)
ESM 2(XLSX 571 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Christina M. Woo
    • 1
  • Alejandra Felix
    • 1
  • Lichao Zhang
    • 2
  • Joshua E. Elias
    • 2
  • Carolyn R. Bertozzi
    • 1
    • 3
  1. 1.Department of ChemistryStanford UniversityStanfordUSA
  2. 2.Chemical and Systems BiologyStanford UniversityStanfordUSA
  3. 3.Howard Hughes Medical InstituteStanford UniversityStanfordUSA

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