title: A novel core fucose-specific lectin from the mushroom Pholiota squarrosa.
abstract: Fucα1-6 oligosaccharide has a variety of biological functions and serves as a biomarker for hepatocellular carcinoma because of the elevated presence of fucosylated α-fetoprotein (AFP) in this type of cancer. In this study we purified a novel Fucα1-6-specific lectin from the mushroom Pholiota squarrosa by ion-exchange chromatography and affinity chromatography on thyroglobulin-agarose. The purified lectin was designated as PhoSL (P. squarrosa lectin). SDS-PAGE, MALDI-TOF mass spectrometry, and N-terminal amino acid sequencing indicate that PhoSL has a molecular mass of 4.5 kDa and consists of 40 amino acids (NH(2)-APVPVTKLVCDGDTYKCTAYLDFGDGRWVAQWDTNVFHTG-OH). Isoelectric focusing of the lectin showed bands near pI 4.0. The lectin activity was stable between pH 2.0 and 11.0 and at temperatures ranging from 0 to 100 °C for incubation times of 30 min. When PhoSL was investigated with frontal affinity chromatography using 132 pyridylaminated oligosaccharides, it was found that the lectin binds only to core α1-6-fucosylated N-glycans and not to other types of fucosylated oligosaccharides, such as α1-2-, α1-3-, and α1-4-fucosylated glycans. Furthermore, PhoSL bound to α1-6-fucosylated AFP but not to non-fucosylated AFP. In addition, PhoSL was able to demonstrate the differential expression of α1-6 fucosylation between primary and metastatic colon cancer tissues. Thus, PhoSL will be a promising tool for analyzing the biological functions of α1-6 fucosylation and evaluating Fucα1-6 oligosaccharides as cancer biomarkers.
authors.affiliation: m the J-Oil Mills, Inc., 11, Kagetoricho, Yokohama, Kanagawa 245-0064, Japan. yuka.kobayashi@j-oil.com
authors.forename: Yuka
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tateno
authors.forename: Hideo
authors.initials: H
authors.lastname: Dohra
authors.forename: Kenta
authors.initials: K
authors.lastname: Moriwaki
authors.forename: Eiji
authors.initials: E
authors.lastname: Miyoshi
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kawagishi
mesh_terms: D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D045744:Cell Line, Tumor; D005643:Fucose / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D037102:Lectins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055368:Pholiota / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D055550:Protein Stability
chemical_list: D000951:Antigens, Neoplasm; D005656:Fungal Proteins; D037102:Lectins; D009844:Oligosaccharides; D005643:Fucose