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High-efficiency labeling of sialylated glycoproteins on living cells

Nature Methods volume 6, pages 207209 (2009) | Download Citation

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.

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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

Author notes

    • Ying Zeng
    •  & T N C Ramya

    These authors contributed equally to this work.

Affiliations

  1. Department of Chemical Physiology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, California 92037, USA.

    • Ying Zeng
    • , T N C Ramya
    •  & James C Paulson
  2. Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, California 92037, USA.

    • Anouk Dirksen
    •  & Philip E Dawson
  3. Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Rd., La Jolla, California 92037, USA.

    • Anouk Dirksen
    •  & Philip E Dawson

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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

Correspondence to James C Paulson.

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    Supplementary Text and Figures

    Supplementary Figures 1–8, Supplementary Methods

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DOI

https://doi.org/10.1038/nmeth.1305

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