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Chemical remodelling of cell surfaces in living animals

Nature volume 430, pages 873–877 (2004)Cite this article

Abstract

Cell surfaces are endowed with biological functionality designed to mediate extracellular communication. The cell-surface repertoire can be expanded to include abiotic functionality through the biosynthetic introduction of unnatural sugars into cellular glycans, a process termed metabolic oligosaccharide engineering1,2. This technique has been exploited in fundamental studies of glycan-dependent cell–cell and virus–cell interactions3,4,5 and also provides an avenue for the chemical remodelling of living cells6,7,8. Unique chemical functional groups can be delivered to cell-surface glycans by metabolism of the corresponding unnatural precursor sugars. These functional groups can then undergo covalent reaction with exogenous agents bearing complementary functionality. The exquisite chemical selectivity required of this process is supplied by the Staudinger ligation of azides and phosphines, a reaction that has been performed on cultured cells without detriment to their physiology7,9. Here we demonstrate that the Staudinger ligation can be executed in living animals, enabling the chemical modification of cells within their native environment. The ability to tag cell-surface glycans in vivo may enable therapeutic targeting and non-invasive imaging of changes in glycosylation during disease progression.

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Figure 1: The Staudinger ligation and metabolic oligosaccharide engineering.
Figure 2: Ac4ManNAz is metabolized in vivo.
Figure 3: Analysis of SiaNAz on cells and in tissues.
Figure 4: The Staudinger ligation proceeds in vivo.

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References

  1. Keppler, O. T., Horstkorte, R., Pawlita, M., Schmidts, C. & Reutter, W. Biochemical engineering of the N-acyl side chain of sialic acid: biological implications. Glycobiology 11, 11R–18R (2001)

    Article  CAS  Google Scholar 

  2. Dube, D. H. & Bertozzi, C. R. Metabolic oligosaccharide engineering as a tool for glycobiology. Curr. Opin. Chem. Biol. 7, 616–625 (2003)

    Article  CAS  Google Scholar 

  3. Buttner, B. et al. Biochemical engineering of cell surface sialic acids stimulates axonal growth. J. Neurosci. 22, 8869–8875 (2002)

    Article  CAS  Google Scholar 

  4. Keppler, O. T. et al. Biosynthetic modulation of sialic acid-dependent virus-receptor interactions of two primate polyoma viruses. J. Biol. Chem. 270, 1308–1314 (1995)

    Article  CAS  Google Scholar 

  5. Charter, N. W., Mahal, L. K., Koshland, D. E. & Bertozzi, C. R. Differential effects of unnatural sialic acids on the polysialylation of the neural cell adhesion molecule and neuronal behavior. J. Biol. Chem. 277, 9255–9261 (2002)

    Article  CAS  Google Scholar 

  6. Mahal, L. K., Yarema, K. J. & Bertozzi, C. R. Engineering chemical reactivity on cell surfaces through oligosaccharide biosynthesis. Science 276, 1125–1128 (1997)

    Article  CAS  Google Scholar 

  7. Saxon, E. & Bertozzi, C. R. Cell surface engineering by a modified Staudinger reaction. Science 287, 2007–2010 (2000)

    Article  ADS  CAS  Google Scholar 

  8. Kayser, H. et al. Biosynthesis of a nonphysiological sialic acid in different rat organs, using N-propanoyl-d-hexosamines as precursors. J. Biol. Chem. 267, 16934–16938 (1992)

    CAS  PubMed  Google Scholar 

  9. Saxon, E. et al. Investigating cellular metabolism of synthetic azidosugars with the Staudinger ligation. J. Am. Chem. Soc. 124, 14893–14902 (2002)

    Article  CAS  Google Scholar 

  10. Hang, H. C., Yu, C., Kato, D. L. & Bertozzi, C. R. A metabolic labeling approach toward proteomic analysis of mucin-type O-linked glycosylation. Proc. Natl Acad. Sci. USA 100, 14846–14851 (2003)

    Article  ADS  CAS  Google Scholar 

  11. Vocadlo, D. J., Hang, H. C., Kim, E. J., Hanover, J. A. & Bertozzi, C. R. A chemical approach for identifying O-GlcNAc-modified proteins in cells. Proc. Natl Acad. Sci. USA 100, 9116–9121 (2003)

    Article  ADS  CAS  Google Scholar 

  12. Kohn, M. & Breinbauer, R. The Staudinger ligation—A gift to chemical biology. Angew. Chem. Int. Edn Engl. 43, 3106–3116 (2004)

    Article  Google Scholar 

  13. Ovaa, H. et al. Chemistry in living cells: Detection of active proteasomes by a two-step labeling strategy. Angew. Chem. Int. Edn Engl. 42, 3626–3629 (2003)

    Article  CAS  Google Scholar 

  14. Kim, Y. J. & Varki, A. Perspectives on the significance of altered glycosylation of glycoproteins in cancer. Glycoconj. J. 14, 569–576 (1997)

    Article  CAS  Google Scholar 

  15. Fukuda, M. Possible roles of tumor-associated carbohydrate antigens. Cancer Res. 56, 2237–2244 (1996)

    CAS  PubMed  Google Scholar 

  16. Renkonen, J., Tynninen, O., Hayry, P., Paavonen, T. & Renkonen, R. Glycosylation might provide endothelial zip codes for organ-specific leukocyte traffic into inflammatory sites. Am. J. Pathol. 161, 543–550 (2002)

    Article  CAS  Google Scholar 

  17. Luchansky, S. J. et al. Constructing azide-labeled cell surfaces using polysaccharide biosynthetic pathways. Methods Enzymol. 362, 249–272 (2003)

    Article  CAS  Google Scholar 

  18. Kavarana, M. J., Kovaleva, E. G., Creighton, D. J., Wollman, M. B. & Eiseman, J. L. Mechanism-based competitive inhibitors of glyoxalase I: Intracellular delivery, in vitro antitumor activities, and stabilities in human serum and mouse serum. J. Med. Chem. 42, 221–228 (1999)

    Article  CAS  Google Scholar 

  19. Morton, C. L. et al. Activation of CPT-11 in mice: Identification and analysis of a highly effective plasma esterase. Cancer Res. 60, 4206–4210 (2000)

    CAS  PubMed  Google Scholar 

  20. Soares, E. R. Identification of a new allele of Es-I segregating in an inbred strain of mice. Biochem. Genet. 17, 577–583 (1979)

    Article  CAS  Google Scholar 

  21. Kiick, K. L., Saxon, E., Tirrell, D. A. & Bertozzi, C. R. Incorporation of azides into recombinant proteins for chemoselective modification by the Staudinger ligation. Proc. Natl Acad. Sci. USA 99, 19–24 (2002)

    Article  ADS  CAS  Google Scholar 

  22. Stasche, R. et al. A bifunctional enzyme catalyzes the first two steps in N-acetylneuraminic acid biosynthesis of rat liver—Molecular cloning and functional expression of UDP-N-acetyl-glucosamine 2-epimerase/N-acetylmannosamine kinase. J. Biol. Chem. 272, 24319–24324 (1997)

    Article  CAS  Google Scholar 

  23. Luchansky, S. J., Argade, S., Hayes, B. K. & Bertozzi, C. R. Metabolic functionalization of recombinant glycoproteins. Biochemistry (in the press)

  24. Shaw, C. F. Gold-based therapeutic agents. Chem. Rev. 99, 2589–2600 (1999)

    Article  CAS  Google Scholar 

  25. Jurisson, S. S. & Lydon, J. D. Potential technetium small molecule radiopharmaceuticals. Chem. Rev. 99, 2205–2218 (1999)

    Article  CAS  Google Scholar 

  26. Codington, J. F., Klein, G., Silber, C., Linsley, K. B. & Jeanloz, R. W. Variations in the sialic acid compositions in glycoproteins of mouse ascites tumor-cell surfaces. Biochemistry 18, 2145–2149 (1979)

    Article  CAS  Google Scholar 

  27. Sell, S. Cancer-associated carbohydrates identified by monoclonal antibodies. Hum. Pathol. 21, 1003–1019 (1990)

    Article  CAS  Google Scholar 

  28. Kolb, H. C. & Sharpless, K. B. The growing impact of click chemistry on drug discovery. Drug Discov. Today 8, 1128–1137 (2003)

    Article  CAS  Google Scholar 

  29. Martin, D. C., Fowlkes, J. L., Babic, B. & Khokha, R. Insulin-like growth factor II signaling in neoplastic proliferation is blocked by transgenic expression of the metalloproteinase inhibitor TIMP-1. J. Cell Biol. 146, 881–892 (1999)

    Article  CAS  Google Scholar 

  30. Reichner, J. S., Whiteheart, S. W. & Hart, G. W. Intracellular trafficking of cell surface sialoglycoconjugates. J. Biol. Chem. 263, 16316–16326 (1988)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank A. Jamieson, S. Luchansky, H. Hang and P. Drake for discussions. J.A.P. was supported by a HHMI Predoctoral Fellowship and D.H.D. was supported by a National Science Foundation Predoctoral Fellowship. Sialic acid analysis was performed by the UCSD GRTC Glycotechnology Core Resource. This work was supported by grants from Johnson & Johnson (Focused Giving Grant), the Mizutani Foundation for Glycoscience, the US Department of Energy and the National Institutes of Health.

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Author notes

  1. Jennifer A. Prescher and Danielle H. Dube: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry, University of California, Berkeley, California, 94720, USA

    Jennifer A. Prescher, Danielle H. Dube & Carolyn R. Bertozzi

  2. Department of Molecular and Cell Biology and Howard Hughes Medical Institute, University of California, Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, 94720, USA

    Carolyn R. Bertozzi

Authors
  1. Jennifer A. Prescher
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Correspondence to Carolyn R. Bertozzi.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure 1

This figure contains a lectin blot depicting the sialoglycoprotein profiles of murine tissue lysates. (DOC 721 kb)

Supplementary Figure 2

This figure contains a Western blot of in vivo Staudinger ligation products on serum glycoproteins. (DOC 1616 kb)

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Prescher, J., Dube, D. & Bertozzi, C. Chemical remodelling of cell surfaces in living animals. Nature 430, 873–877 (2004). https://doi.org/10.1038/nature02791

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