Journal: Nature Communications

Article Title: Intelectin-2 is a broad-spectrum antimicrobial lectin

doi: 10.1038/s41467-025-67099-4

Figure Lengend Snippet: A Overlay of carbohydrate recognition domain of hItln1 (wheat, PDB ID 4WMY) and mItln2 (teal, predicted model), showing calcium (green), conserved calcium coordination residues, and the presence of alanine instead of tryptophan at position 288 in mItln2. B Binding profiles of recombinant mItln2 (25 µg/mL) to microbial (left) and mammalian (right) glycan microarrays from the National Center for Functional Glycomics (NCFG). Graphical representations of top glycan hits from the glycan arrays are included. Complete microarray data are found in Supplementary Data . Data are shown as mean ± SD ( n = 4 technical replicates). RFU, Relative Fluorescence Unit. C Glycan structures bound by mItln2 in glycan arrays. All hits contain either a terminal Gal p or 6SO 3 -Gal p , shown in red. D BLI trace of mItln2 binding to immobilized biotinylated-β-Gal p at varying concentrations, with an apparent Κ d of 1.5 µM. E Dot blot analysis of immobilized MUC2, MUC5AC, and MUC5B, probed with 0.5 μM StrepII-mItln2 in the presence of Ca 2+ or EDTA. F Images of 5 μM StrepII-mItln2 (magenta) binding to 0.01% (w/v) fluorescently labeled MUC5AC (yellow). MUC5AC without lectin treatment served as a control. Scale bars, 20 µm. G Spectroscopic assay for cross-linking of 0.01% (w/v) mucins with varying amounts of mItln2, measured by the increase in absorbance at 405 nm. Schematic of the cross-linking assay is shown on the left. Data are shown as mean ± SD ( n = 3 technical replicates). H Time-lapse images of 0.01% (w/v) fluorescently labeled MUC5AC (yellow) treated with 5 μM mItln2 (magenta). Binding of mItln2 was detected with Strep antibody. Scale bars, 20 µm. I Spectroscopic assay for cross-linking of 0.01% (w/v) lactose-functionalized glycopolymer by mItln2, measured by the increase in absorbance at 405 nm. Data are shown as mean ± SD ( n = 3 technical replicates). Structure of the lactose-functionalized trans-poly (norbornene) is shown on the top. Results in ( D ), ( E ), and ( G ) are representative of three independent experiments. Results in ( F ), ( H ), and ( I ) are representative of two independent experiments. Source data are provided as source data file.

Article Snippet: Lectin binding to mammalian glycans was assessed using the Consortium for Functional Glycomics (CFG) version 5.5 microarray and RayBiotech slides (Cat# GA-Glycan-300-1).

Techniques: Binding Assay, Recombinant, Glycoproteomics, Functional Assay, Microarray, Fluorescence, Dot Blot, Labeling, Control

Journal: Nature Communications

Article Title: Intelectin-2 is a broad-spectrum antimicrobial lectin

doi: 10.1038/s41467-025-67099-4

Figure Lengend Snippet: A Comparison of carbohydrate recognition domain of hItln1 (top, PDB ID 4WMY) and hItln2 (bottom, predicted model), showing conserved calcium coordination residues and a serine replacing tyrosine at position 309 in hItln2. ( B – D ) BLI traces of hItln2 binding to immobilized biotinylated-β-Gal f (a hItln1 ligand) ( B ), biotinylated-β-Gal p ( C ), and biotinylated-6SO 3 LacNAc ( D ). In ( B ) and ( C ), 1.5 μM protein was used. In ( D ), 10 μM protein was used. Data were normalized by background subtraction, using biotin-loaded streptavidin. E Binding of recombinant hItln2 (25 µg/mL) to a microbial glycan microarray from NCFG. Graphical representations of top glycan hits from the glycan arrays are included. Complete microarray data are found in Supplemental Data 2. Data are shown as mean ± SD ( n = 4 technical replicates). F Glycan structures bound by hItln2 in glycan arrays. All hits contain either a Gal p or ethanolamine, shown in red. G Dot blot analysis of MUC2, MUC5AC, and MUC5B, spotted on the membrane and probed with 1 μM hItln2 in the presence of Ca 2+ or EDTA. Binding of hItln2 was detected using a Strep-HRP antibody. H Images of 5 μM hItln2 (magenta) binding to 0.01% (w/v) fluorescently labeled MUC2 (yellow). Binding of hItln2 was detected with Strep antibody. Scale bars, 20 µm. Results in ( B ), ( C ), ( D ) and ( G ) are representative of three independent experiments. Result in ( H ) is representative of two independent experiments. Source data are provided as source data file.

Article Snippet: Lectin binding to mammalian glycans was assessed using the Consortium for Functional Glycomics (CFG) version 5.5 microarray and RayBiotech slides (Cat# GA-Glycan-300-1).

Techniques: Comparison, Binding Assay, Recombinant, Glycoproteomics, Microarray, Dot Blot, Membrane, Labeling