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Chemical remodelling of cell surfaces in living animals

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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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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Correspondence to Carolyn R. Bertozzi.

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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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