Monitoring dynamic glycosylation in vivo using supersensitive click chemistry

Hao Jiang, Tianqing Zheng, Aime Lopez-Aguilar, Lei Feng, Felix Kopp, Florence L. Marlow, Peng Wu

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

To monitor the kinetics of biological processes that take place within the minute time scale, simple and fast analytical methods are required. In this article, we present our discovery of an azide with an internal Cu(I)-chelating motif that enabled the development of the fastest protocol for Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) to date, and its application toward following the dynamic process of glycan biosynthesis. We discovered that an electron-donating picolyl azide boosted the efficiency of the ligand-accelerated CuAAC 20-38-fold in living systems with no apparent toxicity. With a combination of this azide and BTTPS, a tris(triazolylmethyl)amine-based ligand for Cu(I), we were able to detect newly synthesized cell-surface glycans by flow cytometry using as low as 1 nM of a metabolic precursor. This supersensitive chemistry enabled us to monitor the dynamic glycan biosynthesis in mammalian cells and in early zebrafish embryogenesis. In live mammalian cells, we discovered that it takes approximately 30-45 min for a monosaccharide building block to be metabolized and incorporated into cell-surface glycoconjugates. In zebrafish embryos, the labeled glycans could be detected as early as the two-cell stage. To our knowledge, this was the first time that newly synthesized glycans were detected at the cleavage period (0.75-2 hpf) in an animal model using bioorthogonal chemistry.

Original languageEnglish (US)
Pages (from-to)698-706
Number of pages9
JournalBioconjugate Chemistry
Volume25
Issue number4
DOIs
StatePublished - Apr 16 2014

Fingerprint

Click Chemistry
Glycosylation
Biosynthesis
Azides
Polysaccharides
Ligands
Cells
Cycloaddition
Flow cytometry
Monitoring
Chelation
Toxicity
Zebrafish
Amines
Animals
Kinetics
Electrons
Biological Phenomena
Glycoconjugates
Alkynes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Jiang, H., Zheng, T., Lopez-Aguilar, A., Feng, L., Kopp, F., Marlow, F. L., & Wu, P. (2014). Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. Bioconjugate Chemistry, 25(4), 698-706. https://doi.org/10.1021/bc400502d

Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. / Jiang, Hao; Zheng, Tianqing; Lopez-Aguilar, Aime; Feng, Lei; Kopp, Felix; Marlow, Florence L.; Wu, Peng.

In: Bioconjugate Chemistry, Vol. 25, No. 4, 16.04.2014, p. 698-706.

Research output: Contribution to journalArticle

Jiang, H, Zheng, T, Lopez-Aguilar, A, Feng, L, Kopp, F, Marlow, FL & Wu, P 2014, 'Monitoring dynamic glycosylation in vivo using supersensitive click chemistry', Bioconjugate Chemistry, vol. 25, no. 4, pp. 698-706. https://doi.org/10.1021/bc400502d
Jiang H, Zheng T, Lopez-Aguilar A, Feng L, Kopp F, Marlow FL et al. Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. Bioconjugate Chemistry. 2014 Apr 16;25(4):698-706. https://doi.org/10.1021/bc400502d
Jiang, Hao ; Zheng, Tianqing ; Lopez-Aguilar, Aime ; Feng, Lei ; Kopp, Felix ; Marlow, Florence L. ; Wu, Peng. / Monitoring dynamic glycosylation in vivo using supersensitive click chemistry. In: Bioconjugate Chemistry. 2014 ; Vol. 25, No. 4. pp. 698-706.
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