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. 2019 Apr 17;10:1799. doi: 10.1038/s41467-019-09608-w

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© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2019

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Fig. 2 — One-step glycan labeling enabled by recombinant bacterial glycosyltransferases. The Pm2,3ST-M144D, Pd2,6ST, or Hp1,3ST-mediated incorporation of unnatural sugars conjugated to a fluorescent dye (Cy3) or an affinity tag (biotin), enabled a One-step cell-surface glycan labeling. a Nucleotides and analogs functionalized with biotin tag (CMP-SiaNAz-biotin and GDP-FucAz-biotin) or with Cy3 florescent dye (CMP-SiaNAz-Cy3 and GDP-FucAz-Cy3). b Direct STs-catalyzed conjugation of Cy3 (magenta) for imaging of live cell glycans. c Hp1,3FT-catalyzed conjugation of Cy3 (magenta) for imaging of live cell glycans. In b and c, cells were visualized by bright field images and DAPI staining (blue). Scale bar, 20 μm. d Time-dependence of activities of recombinant bacterial and human STs for cell-surface glycan labeling with CMP-SiaNAz-biotin. e Activity of Hp1,3FT using GDP-FucAz-biotin to conjugate biotin onto live cell-surface glycan directly. In d and e, error bars represent the standard deviation of three biological replicates. f, g Enzyme-assisted incorporation of biotin was mainly on N-linked glycans on CHO cells and CHO Lec2 cells, while CHO mutant Lec8 cells without LacNAc were not labeled. Protein loading was depicted by Coomassie blue staining or anti-tubulin western blot. Source data for figures d–g are provided as a Source Data file