Journal Article

New l-Rhamnose-Binding Lectin from the Bivalve Glycymeris yessoensis: Purification, Partial Structural Characterization and Antibacterial Activity

Chikalovets, Irina V., Bulanova, Tatyana A., Filshtein, Alina P., Shilova, Nadezhda V., Rogozhin, Eugene A., Ziganshin, Rustam H., Mizgina, Tatyana O., Molchanova, Valentina I., Chernikov, Oleg V.

2023

Overview

In this study, a new l-rhamnose-binding lectin (GYL-R) from the hemolymph of bivalve Glycymeris yessoensis was purified using affinity and ion-exchange chromatography and functionally characterized. Lectin antimicrobial activity was examined in different ways. The lectin was inhibited by saccharides possessing the same configuration of hydroxyl groups at C-2 and C-4, such as l-rhamnose, d-galactose, lactose, l-arabinose and raffinose. Using the glycan microarray approach, natural carbohydrate ligands were established for GYL-R as l-Rha and glycans containing the α-Gal residue in the terminal position. The GYL-R molecular mass determined by MALDI-TOF mass spectrometry was 30,415 Da. The hemagglutination activity of the lectin was not affected by metal ions. The lectin was stable up to 75 °C and between pH 4.0 and 12.0. The amino acid sequence of the five GYL-R segments was obtained with nano-ESI MS/MS and contained both YGR and DPC-peptide motifs which are conserved in most of the l-rhamnose-binding lectin carbohydrate recognition domains. Circular dichroism confirmed that GYL is a α/β-protein with a predominance of the random coil. Furthermore, GYL-R was able to bind and suppress the growth of the Gram-negative bacteria E. coli by recognizing lipopolysaccharides. Together, these results suggest that GYL-R is a new member of the RBL family which participates in the self-defense mechanism against bacteria and pathogens with a distinct carbohydrate-binding specificity.

• Publisher: MDPI AG
• Subject: Amino acid sequence / Amino acid sequences / Amino acids / Animals / Anti-Bacterial Agents - pharmacology / Antibacterial activity / antibacterial properties / Antibiotics / Antimicrobial activity / Arabinose / Bacteria / Binding / bivalve lectin / Bivalvia / Blood groups / carbohydrate binding / Carbohydrates / Cell cycle / Chromatography / Circular dichroism / circular dichroism spectroscopy / D-Galactose / Defence mechanisms / Dichroism / Disease resistance / DNA / E coli / Eggs / Escherichia coli / Galactose / Glycan / Glycymeris / Gram-negative bacteria / Hemagglutination / Hemolymph / Hydroxyl groups / innate immune / Invertebrates / Ion exchange / Ion-exchange chromatography / l -rhamnose-binding lectin / L-Rhamnose / Lactose / Lectins / Lectins - pharmacology / Ligands / Lipopolysaccharides / Mass spectrometry / Mass spectroscopy / matrix-assisted laser desorption-ionization mass spectrometry / Metal ions / Metals / microarray technology / microorganism binding / Microorganisms / Molecular weight / Mollusks / Nucleotide sequence / Pathogens / Polysaccharides / Proteins / Raffinose / Random coil / Rhamnose / Saccharides / Specificity / Structural analysis / Tandem Mass Spectrometry / Water purification