Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

Enrichment of glycopeptides for glycan structure and attachment site identification

Nature Methods volume 6, pages 809–811 (2009)Cite this article

Abstract

We present a method to enrich for glycoproteins from proteomic samples. Sialylated glycoproteins were selectively periodate-oxidized, captured on hydrazide beads, trypsinized and released by acid hydrolysis of sialic acid glycosidic bonds. Mass spectrometric fragment analysis allowed identification of glycan structures, and additional fragmentation of deglycosylated ions yielded peptide sequence information, which allowed glycan attachment site and protein identification. We identified 36 N-linked and 44 O-linked glycosylation sites on glycoproteins from human cerebrospinal fluid.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Capture-and-release and mass spectrometry analyses of enriched glycopeptides.
Figure 2: Fragment mass spectra of the tryptic O-glycosylated SAAS peptide from ProSAAS.

Similar content being viewed by others

References

  1. Varki, A. et al. Essentials of Glycobiology (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 2009).

  2. Varki, A. Nature 446, 1023–1029 (2007).

    Article  CAS  PubMed  Google Scholar 

  3. Grewal, P.K. et al. Nat. Med. 14, 648–655 (2008).

    Article  CAS  PubMed  Google Scholar 

  4. Ohtsubo, K. & Marth, J.D. Cell 126, 855–867 (2006).

    Article  CAS  PubMed  Google Scholar 

  5. Fujimura, T. et al. Int. J. Cancer 122, 39–49 (2008).

    Article  CAS  PubMed  Google Scholar 

  6. Wada, Y. et al. Glycobiology 17, 411–422 (2007).

    Article  CAS  PubMed  Google Scholar 

  7. Zeng, Y., Ramya, T.N.C., Dirksen, A., Dawson, P.E. & Paulson, J.C. Nat. Methods 6, 207–209 (2009).

    Article  CAS  PubMed  Google Scholar 

  8. Zhang, H., Li, X.J., Martin, D.B. & Aebersold, R. Nat. Biotechnol. 21, 660–666 (2003).

    Article  CAS  PubMed  Google Scholar 

  9. Hwang, H. et al. Mass Spectrom. Rev. advance online publication, doi:10.1002/mas.20221 (8 April 2009).

  10. Satomi, Y., Shimonishi, Y., Hase, T. & Takao, T. Rapid Commun. Mass Spectrom. 18, 2983–2988 (2004).

    Article  CAS  PubMed  Google Scholar 

  11. Sihlbom, C., Davidsson, P., Sjögren, M., Wahlund, L.O. & Nilsson, C.L. Neurochem. Res. 7, 1332–1340 (2008).

    Article  Google Scholar 

  12. Wilson, I.B., Gavel, Y. & von Heijne, G. Biochem. J. 275, 529–534 (1991).

    Article  CAS  PubMed  Google Scholar 

  13. Zhang, Y. et al. J. Biol. Chem. 278, 573–584 (2003).

    Article  CAS  PubMed  Google Scholar 

  14. Alley, W.R., Mechref, Y. & Novotny, M.V. Rapid Commun. Mass Spectrom. 23, 161–170 (2009).

    Article  CAS  PubMed  Google Scholar 

  15. Kubota, K. et al. Anal. Chem. 80, 3693–3698 (2008).

    Article  CAS  PubMed  Google Scholar 

  16. Nilsson, J., Rydell, G.E., Le Pendu, J. & Larson, G. Glycoconj. J advance online publication, doi:10.1007/s10719-009-9237-x (23 April 2009).

  17. Strohalm, M., Hassman, M., Košata, B. & Kodíček, M. Rapid Commun. Mass Spectrom. 22, 905–908 (2008).

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

The work was supported by grants from the Swedish Research Council (8266), governmental grants to the Sahlgrenska University Hospital, Torsten and Ragnar Söderbergs Foundation, the Inga-Britt and Arne Lundberg Research Foundation, Knut and Alice Wallenberg Foundation and cNeupro.

Author information

Authors and Affiliations

  1. Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden

    Jonas Nilsson, Ulla Rüetschi, Adnan Halim, Camilla Hesse & Göran Larson

  2. The Proteomics Core Facility, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

    Elisabet Carlsohn

  3. Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, University of Gothenburg, Sahlgrenska University Hospital, Mölndal, Sweden

    Gunnar Brinkmalm

Authors
  1. Jonas Nilsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ulla Rüetschi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Adnan Halim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Camilla Hesse
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Elisabet Carlsohn
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gunnar Brinkmalm
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Göran Larson
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

J.N. conceived and designed the method, performed experiments, analyzed data and wrote the manuscript; U.R. designed the mass spectrometry setup, performed experiments, analyzed data and wrote the manuscript; A.H. performed experiments, analyzed data and wrote the manuscript; C.H. performed experiments and analyzed data; E.C. and G.B. designed the mass spectrometry setup and performed experiments; and G.L. designed the method and wrote the manuscript.

Corresponding author

Correspondence to Jonas Nilsson.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 and Supplementary Tables 1–3 (PDF 692 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nilsson, J., Rüetschi, U., Halim, A. et al. Enrichment of glycopeptides for glycan structure and attachment site identification. Nat Methods 6, 809–811 (2009). https://doi.org/10.1038/nmeth.1392

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1392

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing