title: Mannose-specific lectin from the mushroom Hygrophorus russula.
abstract: 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, α1-6 mannobiose was the strongest inhibitor and isomaltose, Glcα1-6Glc, was the second strongest one, among mono- and oligosaccharides tested. Frontal affinity chromatography indicated that HRL had the highest affinity for Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc, and non-reducing terminal Manα1-6 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 α1-6 linkage of mannose and glucose, especially the Manα1-6 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.
authors.affiliation: Inovation and Joint Research Center, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Kozue
authors.initials: K
authors.lastname: Sugiyama
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hirai
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ito
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Morita
authors.forename: Hideo
authors.initials: H
authors.lastname: Dohra
authors.forename: Takeomi
authors.initials: T
authors.lastname: Murata
authors.forename: Taichi
authors.initials: T
authors.lastname: Usui
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tateno
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
authors.forename: Yuka
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kawagishi
mesh_terms: D000363:Agaricales / Q000737:chemistry*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D037241:Mannose-Binding Lectins