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A novel core fucose-specific lectin from the mushroom Pholiota squarrosa.
Kobayashi, Yuka; Tateno, Hiroaki; Dohra, Hideo; Moriwaki, Kenta; Miyoshi, Eiji; Hirabayashi, Jun; Kawagishi, Hirokazu.
Afiliação
  • Kobayashi Y; m the J-Oil Mills, Inc., 11, Kagetoricho, Yokohama, Kanagawa 245-0064, Japan. yuka.kobayashi@j-oil.com
J Biol Chem ; 287(41): 33973-82, 2012 Oct 05.
Article em En | MEDLINE | ID: mdl-22872641
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oligossacarídeos / Proteínas Fúngicas / Pholiota / Fucose / Lectinas Limite: Humans Idioma: En Revista: J Biol Chem Ano de publicação: 2012 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Oligossacarídeos / Proteínas Fúngicas / Pholiota / Fucose / Lectinas Limite: Humans Idioma: En Revista: J Biol Chem Ano de publicação: 2012 Tipo de documento: Article País de afiliação: Japão