Fig. 4: Enhancement of programmable glycoprotein tagging by expression of BH-GalNAc-Ts. | Nature Communications

Fig. 4: Enhancement of programmable glycoprotein tagging by expression of BH-GalNAc-Ts.

From: Cell-specific bioorthogonal tagging of glycoproteins

Fig. 4

a Strategy of expanding glycoprotein tagging to include O-GalNAc glycans. Expression of BH-GalNAc-Ts selectively engineered to accommodate bulky chemical tags enhances O-GalNAc tagging in cells expressing NahK/mut-AGX1. b Evaluation of tagging efficiency by feeding transfected K-562 cells with either DMSO, 1 μM Ac4GalN6yne or 2 μM Ac4ManNAlk. Tagging was analysed by in-gel fluorescence and quantification by densitometry. Data are depicted as individual points and means ± SD from at least four independent experiments. c Assessment of tagged glycan subtypes by treating the samples of cells fed with Ac4GalN6yne analysed in (b) with hydrolytic enzymes. Gel regions with lysates of WT- and BH-GalNAc-T1/T2 transfected cells  are scanned independently. Gel images are shown with different intensities to best visualise the effect of enzyme treatment. Data are representative of one out of a total of four replicate labelling experiments performed on two different days. Source data are provided as a Source Data file.

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