maackia amurensis ii lectin mal ii  (Vector Laboratories)

Journal: bioRxiv

Article Title: Sialic acids are a barrier to the entry of non-influenza orthomyxoviruses

doi: 10.64898/2026.01.15.699645

Figure Lengend Snippet: A . Correlations between RNA-seq data for sialyltransferases and infectivity of VSV pseudotypes harbouring WBV, SINUV or OZV GPs, as well as PR8 influenza. Gene expression data were quantified as fragments per kilobase per million (FPKM) and infection rate as the percentage of infected cells. Both metrics were scaled between 0 and 100, transformed as log 2 (x+1), and their Pearson correlation was calculated. Genes for which at least one virus showed a significant ( P < 0.01) correlation are shown. The heatmap depicts the average Pearson correlation from two independent infection assays, as shown in the scale bar. The numbers inside the heatmap represent the levels of significance (1: P < 0.01, 2: P < 0.001, 3: P < 0.0001). VSV pseudotyped with its own glycoprotein was included as a control. B . Examples of scatter plots between infectivity and RNA-seq data for the ST6GALNAC2 and ST3GAL5 genes. Pearson correlation coefficients (r) and their statistical significance ( P ) are shown (ns: not significant). The data points for the SK-MEL-28 cell line are marked in red, and those for IGROV-1 cell line are marked in blue. C . SK-MEL-28 and IGROV-1 cells were incubated with biotin-labelled MAL II and SNA lectins to quantify by flow cytometry α2,3- and α2,6-linked SAs, respectively. Lectin-treated cells were incubated with Streptavidin-Dylight488TM (strep-dye) and the signal emitted by the dye was used to quantify SAs. Left: fluorescence data from two independent experiments. Cells treated without lectin and incubated with the strep-dye (strep-dye only) were used as negative control. A t-test between SK-MEL-28 and IGROV-1 values was carried out using log-transformed data (**: P < 0.01; ***: P < 0.001; ns: P > 0.05). Right: fluorescence level histogram from one representative experiment is shown. Y-axis shows the relative cell count (%) of α2,3- or α2,6-linked SAs-positive cells, and X-axis shows the intensity of fluorescence.

Article Snippet: Biotinylated lectins MAL II ( Maackia Amurensis Lectin II , L-1260-2, Vector labs) and SNA ( Sambucus Nigra Lectin , B-1305-2, Vector labs) that specifically bind to α2,3- and α2,6-SAs, respectively, were added at 4°C for 30 min at a concentration of 20 μg/mL in FACS buffer.

Techniques: RNA Sequencing, Infection, Gene Expression, Transformation Assay, Virus, Control, Incubation, Flow Cytometry, Fluorescence, Negative Control, Cell Characterization

Journal: bioRxiv

Article Title: Sialic acids are a barrier to the entry of non-influenza orthomyxoviruses

doi: 10.64898/2026.01.15.699645

Figure Lengend Snippet: Effect of MALII (α2,3-SA specific) and SNA (α2,6-SA specific) lectins on the infectivity of VSV pseudotypes harbouring WBV, SINUV or OZV GPs, as well as PR8 influenza. VSV pseudotyped with its own glycoprotein was included as a control. Viral entry was measured as the percentage of GFP-positive cells at 18-24 hpi. Three independent assays were carried out for each lectin dose. Right: representative images showing the effects of the combined MALII and SNA treatment at a dose of 100 μg/mL each (scale bar, 1000 µm). Statistical tests comparing the infectivity data at 0 versus 100 μg/mL are provided in the text.

Article Snippet: Biotinylated lectins MAL II ( Maackia Amurensis Lectin II , L-1260-2, Vector labs) and SNA ( Sambucus Nigra Lectin , B-1305-2, Vector labs) that specifically bind to α2,3- and α2,6-SAs, respectively, were added at 4°C for 30 min at a concentration of 20 μg/mL in FACS buffer.

Techniques: Infection, Control

Journal: Nature Communications

Article Title: Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform

doi: 10.1038/s41467-025-62018-z

Figure Lengend Snippet: a Fc-pST3Gal1 (PS1) has four structural motifs. Amino acids shown in cyan (Q108A, Y233A, and Y269F) and magenta (H302A) were mutated in Dead and H302A mutants, respectively. b pST3Gal1 co-crystalized with CMP and Galβ1,3GalNAcα- p NP (PDB: 2WNB). Protein is in green, with selected residues and ligands shown as sticks. CMP is orange, Galβ1,3GalNAcα- p NP is yellow, amino acids mutated in Dead mutant are cyan, and H302A is magenta. c Western blots of Fc-fusion proteins expressed using HEK293T cells under reducing and non-reducing conditions. Detection was performed using anti-human Fc specific IgG. d LC-MS/MS analysis of ST3Gal1 enzymatic activity. Recombinant hST3Gal1 served as positive control. H302A and dead mutants did not yield product (N = 1). e Lectin binding to HEK293T (gray bars), COLO357-FG (green bars) or Calu-3 (blue bars) cells, with or without sialidase treatment, measured using flow cytometry. Cells were incubated with 5 μg/mL fluorescent MALII, PNA, or 1 μg/mL purified Fc-fusion proteins pre-complexed with AF647-conjugated anti-human Fc specific IgG. Data are mean ± STD (N = 3 biological replicates). * p < 0.05 with respect to all other samples; ‡ p < 0.05 with respect to all other samples except samples marked by ‡ are not different from each other. P values calculated using one-way ANOVA followed by Tukey post-test. f Dose dependent binding of PS1 and H302A pre-complexed with AF488-conjugated anti-human Fc specific IgG. HEK293T cells treated with or without sialidase were used. Data are mean (N = 2 biological replicates). Source data including exact P values. Created in BioRender. Neelamegham, S. ( https://BioRender.com/tigz92o ).

Article Snippet: Unconjugated Peanut Agglutinin (PNA, Product code: L-1070), Maackia Amurensis Lectin II (MALII, Product code: L-1260), Erythrina Cristagalli Lectin (ECL, Product code: L-1140), Phaseolus Vulgaris Leucoagglutinin (PHA-L, Product code: L-1110), and H.O.H (Human on Human) Immunodetection kit (Product code: HOH-3000) were from Vector Laboratories (Newark, CA).

Techniques: Mutagenesis, Western Blot, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Recombinant, Positive Control, Binding Assay, Flow Cytometry, Incubation, Purification

Journal: Nature Communications

Article Title: Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform

doi: 10.1038/s41467-025-62018-z

Figure Lengend Snippet: a Schematic of glycan microarray study performed using CFG 5.5 with 562 immobilized glycans. PS1 or H302A were added to microarray at either 5 μg/mL or 50 μg/mL, followed by detection using fluorescent anti-human Fc specific IgG (FL-anti Fc). b , c Lectin binding quantified using relative fluorescence units (RFU) for PS1 (panel b) and H302A (panel c). Data are mean ± STD (N = 6 using six different microarrays). Red arrows with glycan symbol and number represent strong binders. * Glycan 107 is non-specific binder, as its binding is Fc-protein dose independent. d Non-binders of H302A and PS1 are shown. Neither Fc-fusion proteins recognized α2,3-sialyl core-1 O -glycans (219 and 220), α2,6-sialyl core-1 O -glycans (134, 135, 238, and 239), or core-2 O -glycan with α2,3-sialyl LacNAc chains (281, 309, 326, and 560), or a host of N -glycan and glycolipid structures. e HL60 WT cells were cultured with sodium chlorate to prevent sulfation, prior to measuring the binding of 1 μg/mL Fc-fusion protein, pre-complexed with AF647-conjugated anti-human Fc specific IgG. P-selectin Fc binding, but not H302A or PS1 binding, was chlorate sensitive. Results confirm sialylated core-2 O -glycan binding specificity of H302A. Glycan structures are depicted using the Symbol Nomenclature For Glycans standard . Data are mean ± STD (N = 3, biological replicates). P values calculated using two-tailed unpaired Student’s t -test. † p < 0.05 with respect to no sodium chlorate treatment. Source data including exact P values are provided in file. Created in BioRender. Neelamegham, S. ( https://BioRender.com/tigz92o .

Article Snippet: Unconjugated Peanut Agglutinin (PNA, Product code: L-1070), Maackia Amurensis Lectin II (MALII, Product code: L-1260), Erythrina Cristagalli Lectin (ECL, Product code: L-1140), Phaseolus Vulgaris Leucoagglutinin (PHA-L, Product code: L-1110), and H.O.H (Human on Human) Immunodetection kit (Product code: HOH-3000) were from Vector Laboratories (Newark, CA).

Techniques: Glycoproteomics, Microarray, Binding Assay, Fluorescence, Cell Culture, Two Tailed Test

Journal: Nature Communications

Article Title: Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform

doi: 10.1038/s41467-025-62018-z

Figure Lengend Snippet: a Anti-human Fc antibodies were covalently coupled to epoxy-functionalized magnetic beads. The beads were added to HL60 glycoCRISPR library cells that were pre-incubated with PS1 or H302A. Cells binding the Fc-proteins were captured onto magnetic beads, whereas the non-binders remained in solution. These unbound cells were isolated, expanded, and subjected to two more rounds of magnetic enrichment. Genomic DNA was isolated and sequenced to determine sgRNA and corresponding genes regulating Fc-protein binding. b , c Flow cytometry histograms on left side show the HL60 glycoCRISPR library binding profiles for H302A ( b ) and PS1 ( c ) before sorting, after 3rd sort and negative control without Fc-protein. SgRNA enrichment plots on right side identify genes from the 3rd sort, whose depletion by CRISPR-Cas9 also reduced cell-lectin conjugate capture by magnetic beads for H302A ( b ) and PS1 ( c ). Glycogenes critical for lectin binding (FDR < 0.005) were labeled in the figure. Source data are provided in file. Created in BioRender. Neelamegham, S. ( https://BioRender.com/tigz92o

Article Snippet: Unconjugated Peanut Agglutinin (PNA, Product code: L-1070), Maackia Amurensis Lectin II (MALII, Product code: L-1260), Erythrina Cristagalli Lectin (ECL, Product code: L-1140), Phaseolus Vulgaris Leucoagglutinin (PHA-L, Product code: L-1110), and H.O.H (Human on Human) Immunodetection kit (Product code: HOH-3000) were from Vector Laboratories (Newark, CA).

Techniques: Magnetic Beads, Incubation, Binding Assay, Isolation, Protein Binding, Flow Cytometry, Negative Control, CRISPR, Labeling

Journal: Nature Communications

Article Title: Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform

doi: 10.1038/s41467-025-62018-z

Figure Lengend Snippet: a , b Fusing the cytoplasmic and transmembrane domain of type-II transmembrane protein DPP4 at the N-terminus of Fc-proteins (H302A in this example) enabled a mammalian surface display platform. Amino acids mutated to create Lib1 and Lib2 were shown in cyan and magenta, respectively in ( a ). c , d HEK293T WT cells were transfected to surface display TM-PS1, TM-H302A and TM-Dead. Cells were treated with sialidase to eliminate cis -interactions ( c ). AF647-conjugated goat anti-human Fc-specific IgG was used to monitor Fc expression level. Cells with high Fc expression were gated and used in the histogram in panel d as a measure of lectin binding with sialyl core-2 PAA-FITC. e Amino acid residues mutated in Lib1 and Lib2 are shown in Alphafold model of pST3Gal1 (green) and its crystal structure (PDB: 2WNB, gray) co-crystalized with CMP (purple stick) and Galβ1,3GalNAcα- p NP (yellow stick). These residues, which are represented as cyan (Lib1) and magenta (Lib2) sticks, were mutated for optimizing the glycan binding properties of H302A. Neu5Ac superimposed from 5FRE was shown as orange sticks. f Sialyl core-2 PAA-FITC binding to HEK293T SLC35A1 -KO cells transduced to express TM-H302A, Lib1, and Lib2. g Identification of top-6 mutants in Lib2 that are superior binders of sialyl core-2 PAA-FITC. Source data are provided in file. Created in BioRender. Neelamegham, S. ( https://BioRender.com/tigz92o ).

Article Snippet: Unconjugated Peanut Agglutinin (PNA, Product code: L-1070), Maackia Amurensis Lectin II (MALII, Product code: L-1260), Erythrina Cristagalli Lectin (ECL, Product code: L-1140), Phaseolus Vulgaris Leucoagglutinin (PHA-L, Product code: L-1110), and H.O.H (Human on Human) Immunodetection kit (Product code: HOH-3000) were from Vector Laboratories (Newark, CA).

Techniques: Transfection, Expressing, Binding Assay, Glycoproteomics

Journal: Nature Communications

Article Title: Engineering glycosyltransferases into glycan binding proteins using a mammalian surface display platform

doi: 10.1038/s41467-025-62018-z

Figure Lengend Snippet: a tSNE plots showing the binding of PNA, MALII and sCore2 to different human blood cell types, immunoprofiled using 23 antibodies. Representative data are shown for a single donor. Legends show numbers and color labels for each cell population in tSNE plot. b Heatmap showing the average normalized binding of each lectin to different human blood cell types, based on studies with two donors. Heatmaps are arranged based on cell lineage. Some cell types from heatmap do not appear in tSNE plots as these are rare populations, with small number of cells. c Human normal tissue microarray and breast cancer tissue analysis using sCore2. Staining of selected cores are shown, along with higher magnification tissue sections. Data for negative controls using Dead Fc-protein and upon sialidase treatment are presented in Supplementary Information. Results show cell specific staining properties of sCore2. Images are representative of two independent experiments. Main figure scale bar = 400 μm (except 300 μm for breast tumors). Magnified image scale bar = 100 μm. Source data are provided in file.

Article Snippet: Unconjugated Peanut Agglutinin (PNA, Product code: L-1070), Maackia Amurensis Lectin II (MALII, Product code: L-1260), Erythrina Cristagalli Lectin (ECL, Product code: L-1140), Phaseolus Vulgaris Leucoagglutinin (PHA-L, Product code: L-1110), and H.O.H (Human on Human) Immunodetection kit (Product code: HOH-3000) were from Vector Laboratories (Newark, CA).

Techniques: Binding Assay, Microarray, Staining

Journal: The Journal of Biological Chemistry

Article Title: Maackia amurensis seed lectin structure and sequence comparison with other M. amurensis lectins

doi: 10.1016/j.jbc.2025.108466

Figure Lengend Snippet: Amino acid sequence of dimerizing and nondimerizing Maackia amurensis lectins. Alignment of MASL (Sentrimed Maackia amurensis seed lectin) and MAL2 (Vector Labs L-1260-2 based on Yin et al. ) with MAA (EY Laboratories L-7801-1 based on Yamamoto et al. ) with sequences identified by LC-MS/MS in bold font . Signal peptides are italicized , and a cysteine or serine found near the carboxy tail of dimerizing and nondimerizing lectins, respectively, is indicated by a hashtag . A tyrosine at amino acid 250 in MASL and MAA, substituted with alanine in MAL2 is indicated by an exclamation mark . Glycosylation sites identified in MASL are indicated by an italicized dollar sign . Conserved residues, substitutions, and deletions are indicated by asterisks , periods , and dashes , respectively, with amino acids numbered as indicated.

Article Snippet: Alignment of MASL (Sentrimed Maackia amurensis seed lectin) and MAL2 (Vector Labs L-1260-2 based on Yin et al. ( )) with MAA (EY Laboratories L-7801-1 based on Yamamoto et al. ( )) with sequences identified by LC-MS/MS in bold font .

Techniques: Sequencing, Plasmid Preparation, Liquid Chromatography with Mass Spectroscopy, Glycoproteomics

Journal: The Journal of Biological Chemistry

Article Title: Maackia amurensis seed lectin structure and sequence comparison with other M. amurensis lectins

doi: 10.1016/j.jbc.2025.108466

Figure Lengend Snippet: Maackia amurensis lectins resolved by SDS-PAGE. A , MAA, MASL, and MAL2 (15 ug per lane) were resolved by nonreducing and reducing 12% SDS-PAGE and visualized by Coomassie staining along with molecular weight markers as indicated. B , MAA, MASL, and MAL2 (1 ug per lane) were resolved by 12% SDS-PAGE with migration of molecular weight markers as indicated. Lectins were detected by Western blotting with two independent rabbit antibodies as indicated. C , membranes were stained with India ink after Western blotting. MASL, Maackia amurensis seed lectin.

Article Snippet: Alignment of MASL (Sentrimed Maackia amurensis seed lectin) and MAL2 (Vector Labs L-1260-2 based on Yin et al. ( )) with MAA (EY Laboratories L-7801-1 based on Yamamoto et al. ( )) with sequences identified by LC-MS/MS in bold font .

Techniques: SDS Page, Staining, Molecular Weight, Migration, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Maackia amurensis seed lectin structure and sequence comparison with other M. amurensis lectins

doi: 10.1016/j.jbc.2025.108466

Figure Lengend Snippet: Comparison of MASL, MAA, and MAL2 binding to glycans. Lectins were hybridized to glycan arrays to compare target binding affinities. Signal intensities were normalized to positive controls with background subtracted and are shown as mean + SEM (n = 3). Twenty glycans with the highest affinity for MASL are shown. MASL, Maackia amurensis seed lectin.

Article Snippet: Alignment of MASL (Sentrimed Maackia amurensis seed lectin) and MAL2 (Vector Labs L-1260-2 based on Yin et al. ( )) with MAA (EY Laboratories L-7801-1 based on Yamamoto et al. ( )) with sequences identified by LC-MS/MS in bold font .

Techniques: Comparison, Binding Assay, Glycoproteomics