Mannose-specific lectin from the mushroom Hygrophorus russula

作者：Suzuki Tomohiro; Sugiyama Kozue; Hirai Hirofumi; Ito Hiroyuki; Morita Tatsuya; Dohra Hideo; Murata Takeomi; Usui Taichi; Tateno Hiroaki; Hirabayashi Jun; Kobayashi Yuka^*; Kawagishi Hirokazu

来源：Glycobiology, 2012, 22(5): 616-629.

DOI：10.1093/glycob/cwr187

• fungal lectin
• Hygrophorus russula
• mannose-binding lectin
• mushroom

A lectin was purified from the mushroom Hygrophorus russula by affinity chromatography on a Sephadex G-50 column and BioAssist S cation exchange chromatography and designated H. russula lectin (HRL). The results of sodium dodecyl sulfate-polyaclylamidegel electrophoresis, gel filtration and matrix-assisted laser desorption ionization time-of-flight mass spectrometry of HRL indicated that it was composed of four identical 18.5 kDa subunits with no S-S linkage. Isoelectric focusing of the lectin showed bands near pI 6.40. The complete sequence of 175 amino acid residues was determined by amino acid sequencing of intact or enzyme-digested HRL. The sequence showed homology with Grifola frondosa lectin. The cDNA of HRL was cloned from RNA extracted from the mushroom. The open reading frame of the cDNA consisted of 528 bp encoding 176 amino acids. In hemagglutination inhibition assay, alpha 1-6 mannobiose was the strongest inhibitor and isomaltose, Glc alpha 1-6Glc, was the second strongest one, among mono- and oligosaccharides tested. Frontal affinity chromatography indicated that HRL had the highest affinity for Man alpha 1-6(Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4GlcNAc, and non-reducing terminal Man alpha 1-6 was essential for the binding of HRL to carbohydrate chains. The sugar-binding specificity of HRL was also analyzed by using BIAcore. The result from the analysis exhibited positive correlations with that of the hemagglutination inhibition assay. All the results suggested that HRL recognized the alpha 1-6 linkage of mannose and glucose, especially the Man alpha 1-6 bond. HRL showed a mitogenic activity against spleen lymph cells of an F344 rat. Furthermore, an enzyme-linked immunosorbent assay showed strong binding of HRL to human immunodeficiency virus type-1 gp120.

• 出版日期2012-5