Purification, characterization, and cDNA cloning of α-N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera

Mari Kakiuchi, Nozomu Okino, Noriyuki Sueyoshi, Sachiyo Ichinose, Akira Omori, Shun-Ichiro Kawabata, Kuniko Yamaguchi, Makoto Ito

研究成果: ジャーナルへの寄稿記事

22 引用 (Scopus)

抄録

We report here the purification, characterization, and cDNA cloning of a novel N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. The purified lectin showed 19-kDa, 41-kDa, and 60-kDa protein bands on SDS-PAGE, possibly corresponding to a monomer, homodimer, and homotrimer. Interestingly, on 4-20% native PAGE the lectin showed at least nine protein bands, among which oligomers containing six to nine subunits had potent hemagglutination activity for sheep erythrocytes. The hemagglutination activity of the lectin was specifically inhibited by N-acetylgalactosamine, Tn antigen, and blood group A trisaccharide, but not by N-acetylglucosamine, galactose, galactosamine, or blood group B trisaccharide. The specificity of the lectin was further examined using various glycosphingolipids and biotin-labeled lectin. The lectin was found to bind to Gb5Cer, but not Gb4Cer, Gb3Cer, GM1a, GM2, or asialo-GM2, indicating that the lectin specifically binds to the terminal α-GalNAc at the nonreducing end. The hemagglutination activity of the lectin was completely abolished by chelation with EDTA or EGTA and completely restored by the addition of CaCl2. cDNA cloning of the lectin showed that the protein is composed of 168 amino acids, including a signal sequence of 18 residues, and possesses the typical C-type lectin motif. These findings indicate that the protein is a C-type lectin. The recombinant lectin, produced in a soluble form by Escherichia coli, showed binding activity for asialomucin in the presence of Ca2+ but no hemagglutination.

元の言語英語
ページ(範囲)85-94
ページ数10
ジャーナルGlycobiology
12
発行部数2
DOI
出版物ステータス出版済み - 1 1 2002

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Asterina
Starfish
Acetylgalactosamine
Cloning
Lectins
Purification
Organism Cloning
Complementary DNA
Hemagglutination
C-Type Lectins
Proteins
Native Polyacrylamide Gel Electrophoresis
Glycosphingolipids
Galactosamine
Acetylglucosamine
Egtazic Acid
Biotin
Protein Sorting Signals
Chelation
Galactose

All Science Journal Classification (ASJC) codes

  • Biochemistry

これを引用

Purification, characterization, and cDNA cloning of α-N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. / Kakiuchi, Mari; Okino, Nozomu; Sueyoshi, Noriyuki; Ichinose, Sachiyo; Omori, Akira; Kawabata, Shun-Ichiro; Yamaguchi, Kuniko; Ito, Makoto.

:: Glycobiology, 巻 12, 番号 2, 01.01.2002, p. 85-94.

研究成果: ジャーナルへの寄稿記事

Kakiuchi, Mari ; Okino, Nozomu ; Sueyoshi, Noriyuki ; Ichinose, Sachiyo ; Omori, Akira ; Kawabata, Shun-Ichiro ; Yamaguchi, Kuniko ; Ito, Makoto. / Purification, characterization, and cDNA cloning of α-N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. :: Glycobiology. 2002 ; 巻 12, 番号 2. pp. 85-94.
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abstract = "We report here the purification, characterization, and cDNA cloning of a novel N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. The purified lectin showed 19-kDa, 41-kDa, and 60-kDa protein bands on SDS-PAGE, possibly corresponding to a monomer, homodimer, and homotrimer. Interestingly, on 4-20{\%} native PAGE the lectin showed at least nine protein bands, among which oligomers containing six to nine subunits had potent hemagglutination activity for sheep erythrocytes. The hemagglutination activity of the lectin was specifically inhibited by N-acetylgalactosamine, Tn antigen, and blood group A trisaccharide, but not by N-acetylglucosamine, galactose, galactosamine, or blood group B trisaccharide. The specificity of the lectin was further examined using various glycosphingolipids and biotin-labeled lectin. The lectin was found to bind to Gb5Cer, but not Gb4Cer, Gb3Cer, GM1a, GM2, or asialo-GM2, indicating that the lectin specifically binds to the terminal α-GalNAc at the nonreducing end. The hemagglutination activity of the lectin was completely abolished by chelation with EDTA or EGTA and completely restored by the addition of CaCl2. cDNA cloning of the lectin showed that the protein is composed of 168 amino acids, including a signal sequence of 18 residues, and possesses the typical C-type lectin motif. These findings indicate that the protein is a C-type lectin. The recombinant lectin, produced in a soluble form by Escherichia coli, showed binding activity for asialomucin in the presence of Ca2+ but no hemagglutination.",
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T1 - Purification, characterization, and cDNA cloning of α-N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera

AU - Kakiuchi, Mari

AU - Okino, Nozomu

AU - Sueyoshi, Noriyuki

AU - Ichinose, Sachiyo

AU - Omori, Akira

AU - Kawabata, Shun-Ichiro

AU - Yamaguchi, Kuniko

AU - Ito, Makoto

PY - 2002/1/1

Y1 - 2002/1/1

N2 - We report here the purification, characterization, and cDNA cloning of a novel N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. The purified lectin showed 19-kDa, 41-kDa, and 60-kDa protein bands on SDS-PAGE, possibly corresponding to a monomer, homodimer, and homotrimer. Interestingly, on 4-20% native PAGE the lectin showed at least nine protein bands, among which oligomers containing six to nine subunits had potent hemagglutination activity for sheep erythrocytes. The hemagglutination activity of the lectin was specifically inhibited by N-acetylgalactosamine, Tn antigen, and blood group A trisaccharide, but not by N-acetylglucosamine, galactose, galactosamine, or blood group B trisaccharide. The specificity of the lectin was further examined using various glycosphingolipids and biotin-labeled lectin. The lectin was found to bind to Gb5Cer, but not Gb4Cer, Gb3Cer, GM1a, GM2, or asialo-GM2, indicating that the lectin specifically binds to the terminal α-GalNAc at the nonreducing end. The hemagglutination activity of the lectin was completely abolished by chelation with EDTA or EGTA and completely restored by the addition of CaCl2. cDNA cloning of the lectin showed that the protein is composed of 168 amino acids, including a signal sequence of 18 residues, and possesses the typical C-type lectin motif. These findings indicate that the protein is a C-type lectin. The recombinant lectin, produced in a soluble form by Escherichia coli, showed binding activity for asialomucin in the presence of Ca2+ but no hemagglutination.

AB - We report here the purification, characterization, and cDNA cloning of a novel N-acetylgalactosamine-specific lectin from starfish, Asterina pectinifera. The purified lectin showed 19-kDa, 41-kDa, and 60-kDa protein bands on SDS-PAGE, possibly corresponding to a monomer, homodimer, and homotrimer. Interestingly, on 4-20% native PAGE the lectin showed at least nine protein bands, among which oligomers containing six to nine subunits had potent hemagglutination activity for sheep erythrocytes. The hemagglutination activity of the lectin was specifically inhibited by N-acetylgalactosamine, Tn antigen, and blood group A trisaccharide, but not by N-acetylglucosamine, galactose, galactosamine, or blood group B trisaccharide. The specificity of the lectin was further examined using various glycosphingolipids and biotin-labeled lectin. The lectin was found to bind to Gb5Cer, but not Gb4Cer, Gb3Cer, GM1a, GM2, or asialo-GM2, indicating that the lectin specifically binds to the terminal α-GalNAc at the nonreducing end. The hemagglutination activity of the lectin was completely abolished by chelation with EDTA or EGTA and completely restored by the addition of CaCl2. cDNA cloning of the lectin showed that the protein is composed of 168 amino acids, including a signal sequence of 18 residues, and possesses the typical C-type lectin motif. These findings indicate that the protein is a C-type lectin. The recombinant lectin, produced in a soluble form by Escherichia coli, showed binding activity for asialomucin in the presence of Ca2+ but no hemagglutination.

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U2 - 10.1093/glycob/12.2.85

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SN - 0959-6658

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