Fig. 3: GRAB validation by ITC and glycan array analysis.
From: Mutant glycosidases for labeling sialoglycans with high specificity and affinity
GRAB-Sia was subjected to ITC analysis with 3′SL (a) and 6′SL (b) as the ligand. GRAB-Sia3 was subjected to ITC analysis with 3′SL (c) and 6′SL (d) as the ligand. In (a–d), the ITC titration and binding curves are shown. e Monobiotinylated GRABs generated using the BirA-AviTag system were mixed with streptavidin conjugates to yield tetra-GRABs. Created in BioRender. Liang, S. (2025) https://BioRender.com/g78q224. Binding of tetra-GRAB-Sia (f) and tetra-GRAB-Sia3 (g) to different glycan structures was evaluated by glycan array analysis. The bar chart showed relative fluorescence units (RFU) of the 166 glycans by their order of glycan ID on the x-axis. Data are presented as mean ± s.d. of four replicates. Glycan structures are grouped based on the terminal monosaccharide. h Representative glycan substrates of tetra-GRAB-Sia and tetra-GRAB-Sia3. Source data are provided as a Source Data file. The source data for (f, g) are provided in a separate Excel file as Supplementary Data 1.
