Analytical and Bioanalytical Chemistry

, Volume 409, Issue 2, pp 579–588

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

DOI: 10.1007/s00216-016-9934-9

Cite this article as:
Woo, C.M., Felix, A., Zhang, L. et al. Anal Bioanal Chem (2017) 409: 579. doi:10.1007/s00216-016-9934-9
Part of the following topical collections:
  1. Glycomics, Glycoproteomics and Allied Topics


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.


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)

Funding information

Funder NameGrant NumberFunding Note
Bill and Melinda Gates Foundation
    Stanford Undergraduate Advising and Research Student Grant
      Jane Coffin Childs Memorial Fund for Medical Research
        Burroughs Wellcome Fund
          W.M. Keck Foundation Medical Research Program
            National Cancer Institute
            • CA200423
            Howard Hughes Medical Institute

              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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