Metabolic labeling of fucosylated glycans in developing zebrafish.

Karen W. Dehnert, Brendan J. Beahm, Thinh T. Huynh, Jeremy M. Baskin, Scott T. Laughlin, Wei Wang, Peng Wu, Sharon L. Amacher, Carolyn R. Bertozzi

Research output: Contribution to journalArticle

Abstract

Many developmental processes depend on proper fucosylation, but this post-translational modification is difficult to monitor in vivo. Here we applied a chemical reporter strategy to visualize fucosylated glycans in developing zebrafish. Using azide-derivatized analogues of fucose, we metabolically labeled cell-surface glycans and then detected the incorporated azides via copper-free click chemistry with a difluorinated cyclooctyne probe. We found that the fucose salvage pathway enzymes are expressed during zebrafish embryogenesis but that they process the azide-modified substrates inefficiently. We were able to bypass the salvage pathway by using an azide-functionalized analogue of GDP-fucose. This nucleotide sugar was readily accepted by fucosyltransferases and provided robust cell-surface labeling of fucosylated glycans, as determined by flow cytometry and confocal microscopy analysis. We used this technique to image fucosylated glycans in the enveloping layer of zebrafish embryos during the first 5 days of development. This work provides a method to study the biosynthesis of fucosylated glycans in vivo.

Original languageEnglish (US)
Pages (from-to)547-552
Number of pages6
JournalACS Chemical Biology
Volume6
Issue number6
StatePublished - Jun 17 2011
Externally publishedYes

Fingerprint

Zebrafish
Azides
Labeling
Polysaccharides
Salvaging
Fucose
Guanosine Diphosphate Fucose
Click Chemistry
Fucosyltransferases
Flow cytometry
Confocal microscopy
Biosynthesis
Post Translational Protein Processing
Confocal Microscopy
Sugars
Embryonic Development
Copper
Flow Cytometry
Embryonic Structures
Nucleotides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Dehnert, K. W., Beahm, B. J., Huynh, T. T., Baskin, J. M., Laughlin, S. T., Wang, W., ... Bertozzi, C. R. (2011). Metabolic labeling of fucosylated glycans in developing zebrafish. ACS Chemical Biology, 6(6), 547-552.

Metabolic labeling of fucosylated glycans in developing zebrafish. / Dehnert, Karen W.; Beahm, Brendan J.; Huynh, Thinh T.; Baskin, Jeremy M.; Laughlin, Scott T.; Wang, Wei; Wu, Peng; Amacher, Sharon L.; Bertozzi, Carolyn R.

In: ACS Chemical Biology, Vol. 6, No. 6, 17.06.2011, p. 547-552.

Research output: Contribution to journalArticle

Dehnert, KW, Beahm, BJ, Huynh, TT, Baskin, JM, Laughlin, ST, Wang, W, Wu, P, Amacher, SL & Bertozzi, CR 2011, 'Metabolic labeling of fucosylated glycans in developing zebrafish.', ACS Chemical Biology, vol. 6, no. 6, pp. 547-552.
Dehnert KW, Beahm BJ, Huynh TT, Baskin JM, Laughlin ST, Wang W et al. Metabolic labeling of fucosylated glycans in developing zebrafish. ACS Chemical Biology. 2011 Jun 17;6(6):547-552.
Dehnert, Karen W. ; Beahm, Brendan J. ; Huynh, Thinh T. ; Baskin, Jeremy M. ; Laughlin, Scott T. ; Wang, Wei ; Wu, Peng ; Amacher, Sharon L. ; Bertozzi, Carolyn R. / Metabolic labeling of fucosylated glycans in developing zebrafish. In: ACS Chemical Biology. 2011 ; Vol. 6, No. 6. pp. 547-552.
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