Abstract
We describe a simple method for efficiently labeling cell-surface sialic acid–containing glycans on living animal cells. The method uses mild periodate oxidation to generate an aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag. Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins while maintaining high cell viability.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Bond, M.R. & Kohler, J.J. Curr. Opin. Chem. Biol. 11, 52–58 (2007).
Laughlin, S.T., Baskin, J.M., Amacher, S.L. & Bertozzi, C.R. Science 320, 664–667 (2008).
Hangauer, M.J. & Bertozzi, C.R. Angew. Chem. Int. Ed. Engl. 47, 2394–2397 (2008).
Hsu, T.L. et al. Proc. Natl. Acad. Sci. USA 104, 2614–2619 (2007).
Rodionov, V.O., Fokin, V.V. & Finn, M.G. Angew. Chem. Int. Ed. Engl. 44, 2210–2215 (2005).
Nauman, D.A. & Bertozzi, C.R. Biochim. Biophys. Acta 1568, 147–154 (2001).
De Bank, P.A., Kellam, B., Kendall, D.A. & Shakesheff, K.M. Biotechnol. Bioeng. 81, 800–808 (2003).
Gahmberg, C.G. & Andersson, L.C. J. Biol. Chem. 252, 5888–5894 (1977).
Cordes, E.H. & Jencks, W.P. J. Am. Chem. Soc. 84, 826–831 (1962).
Dirksen, A. & Dawson, P.E. Bioconjug. Chem. 19, 2543–2548 (2008).
Dirksen, A., Dirksen, S., Hackeng, T.M. & Dawson, P.E. J. Am. Chem. Soc. 128, 15602–15603 (2006).
Dirksen, A., Hackeng, T.M. & Dawson, P.E. Angew. Chem. Int. Ed. Engl. 45, 7581–7584 (2006).
Kalia, J. & Raines, R.T. Angew. Chem. Int. Ed. Engl. 47, 7523–7526 (2008).
Bhavanandan, V.P., Murray, M. & Davidson, E.A. Glycoconj. J. 5, 467–484 (1988).
Oetke, C. et al. Eur. J. Biochem. 268, 4553–4561 (2001).
Acknowledgements
We thank M. Pawlita (University of Berlin) for providing the BJA-B K88 and K20 cell lines, M. Krieger (Massachusetts Institute of Technology) for providing the CHO ldlD cells, dearly departed Karl Schmidt for the gift of α1-acid glycoprotein, S. Han for assistance with the early experiments and A. Tran-Crie for her expert assistance in manuscript preparation. This work was funded by the US National Institutes of Health grants GM60938 and AI50143 to J.C.P. and GM059380 to P.E.D.
Author information
Authors and Affiliations
Contributions
P.E.D., A.D. and J.C.P. conceived of the method for PAL on cells. A.D. and Y.Z. performed preliminary experiments. Y.Z., T.N.C.R. and J.C.P. designed the reported experiments. Y.Z. and T.N.C.R. performed the majority of the experiments. Y.Z. and T.N.C.R. wrote the manuscript. All authors participated in the analysis of the data and edited the manuscript.
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–8, Supplementary Methods (PDF 1728 kb)
Rights and permissions
About this article
Cite this article
Zeng, Y., Ramya, T., Dirksen, A. et al. High-efficiency labeling of sialylated glycoproteins on living cells. Nat Methods 6, 207–209 (2009). https://doi.org/10.1038/nmeth.1305
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nmeth.1305
This article is cited by
-
Glycoproteomics
Nature Reviews Methods Primers (2022)
-
Bioorthogonal chemistry
Nature Reviews Methods Primers (2021)
-
Cell surface thermal proteome profiling tracks perturbations and drug targets on the plasma membrane
Nature Methods (2021)
-
Selective N-glycan editing on living cell surfaces to probe glycoconjugate function
Nature Chemical Biology (2020)
-
Assembling custom side chains on proteoglycans to interrogate their function in living cells
Nature Communications (2020)