Fig. 2: Glycan binding activity of GnT-IVa lectin domain. | Communications Biology

Fig. 2: Glycan binding activity of GnT-IVa lectin domain.

From: Discovery of a lectin domain that regulates enzyme activity in mouse N-acetylglucosaminyltransferase-IVa (MGAT4A)

Fig. 2

a GST-GnT-IVa lectin domain (#5) was purified from E. coli, and GST tag was cleaved and eliminated. Purity was checked by SDS-PAGE and subsequent CBB staining (left). Elution profiles of the gel filtration analysis during the purification step are shown (right). The same gel filtration analysis was performed in the presence of 2 mM Glc (green) or GlcNAc (red). b Solution NMR analysis of the interaction between 15N-labeled GnT-IVa lectin domain and GlcNAc monosaccharide. Overlayed 1H-15N HSQC spectra of 0.4 mM [15N]GnT-IVa lectin domain at protein-to-ligand ratio of 1:0, 1:1, 1:2, 1:3, 1:5 and 1:7 for GlcNAc. Peak A was used for calculation of dissociation constant. c Solution NMR analysis of the interaction between 15N-labeled GnT-IVa lectin domain and GlcNAcβ1-2Man disaccharide. Overlayed 1H-15N HSQC spectra of 0.4 mM [15N]GnT-IVa lectin domain at protein-to-ligand ratio of 1:0, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5 and 1:3 for GlcNAcβ1-2Man. d Plot of 1H chemical shift (peak A) against ligand-to-protein molar ratio for GlcNAc (filled square) and GlcNAcβ1-2Man (filled circle). e Binding intensity of 157 glycans toward immobilized GnT-IVa lectin domain in frontal affinity chromatography.

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