High-mannose N-glycan-specific lectin from the red alga Kappaphycus striatum (Carrageenophyte) |
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Authors: | Le Dinh Hung Yuichiro Sato Kanji Hori |
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Institution: | a Graduate School of Biosphere Science, Hiroshima University, Kagamiyama 1-4-4, Higashi - Hiroshima 739-8528, Japan b Nhatrang Institute of Technology Research and Application, 2A-Hungvuong Street, Nhatrang City, Khanhhoa Province, Viet Nam c Faculty of Pharmacy, Yasuda Women’s University, 6-13-1 Yasuhigashi, Asaminami, Hiroshima 731-0153, Japan |
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Abstract: | From a fresh sample (1 kg) of cultivated red alga Kappaphycus striatum, three isolectins, KSA-1 (15.1 mg), KSA-2 (58.0 mg) and KSA-3 (6.9 mg), were isolated by a combination of extraction with aqueous ethanol, ethanol precipitation, and ion exchange chromatography. Isolated KSAs were monomeric proteins of about 28 kDa having identical 20 N-terminal amino acid sequences to each other. Their hemagglutination activities were not inhibited by monosaccharides, but inhibited by glycoproteins bearing high-mannose N-glycans. In a binding experiment with pyridylaminated oligosaccharides by centrifugal ultrafiltration-HPLC assay, the isolectin KSA-2 was exclusively bound to high-mannose type N-glycans, but not to other glycans. Including complex types and a pentasaccharide core of N-glycans, indicating that it recognized branched oligomannosides. The binding activity of KSA-2 was slightly different among high-mannose N-glycans examined, indicating that the lectin has a higher affinity for those having the exposed (α1-3) Man in the D2 arm. On the other hand, KSA-2 did not bind to a free oligomannose that is a constituent of the branched oligomannosides, implying that the portion of the core GlcNAc residue(s) of the N-glycans is also essential for binding. Thus, KSA-2 appears to recognize the extended carbohydrate structure with a minimal length of a tetrasaccharide, Man(α1-3)Man(α1-6)Man(β1-4)GlcNAc. This study indicates that K. striatum, which has extensively been cultivated as a source of carrageenan, is a good source of a valuable lectin(s) that is strictly specific for high-mannose N-glycans. |
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Keywords: | CRLL Cycas revoluta leaf lectin CV-N Nostoc ellipsosporum lectin ESA Eucehuma serra lectin ESI-MS electron spray ionization-mass spectrometry GlcNAc d-glucosamine" target="_blank">N-acetyl-d-glucosamine GRFT Griffithsia sp lectin HIV human immunodeficiency virus HPLC high performance liquid chromatography HU hemagglutination unit (hemagglutination titer) KAA Kappaphycus alvarezii lectin KSA Kappaphycus striatum lectin Man mannose MVL Microcystis viridis lectin OAA Oscillatoria agardhii agglutinin PA pyridylaminated PE pyridylethylated SDS sodium dodecyl sulfate SVN Scytonema varium lectin TFA trifluoroacetic acid |
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