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1988 Fiscal Year Final Research Report Summary

Studies on the structure and function of the liver mannan-binding protein by utilizing gene recombinant technology.

Research Project

Project/Area Number 61480429
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionKyoto University

Principal Investigator

KAWASAKI Toshisuke  Faculty of Pharmaceutical Sciences, Kyoto University, 薬学部, 助教授 (50025706)

Co-Investigator(Kenkyū-buntansha) SUGAHARA Kazuyuki  Faculty of Pharmaceutical Sciences, Kyoto University, 薬学部, 助手 (60154449)
Project Period (FY) 1986 – 1988
KeywordsAnimal lectin / primary structure / cDNA / signal peptide / collagen / endogenous ligand / serum glycoprotein / リソソーム酵素
Research Abstract

The function and structure of the liver mannan-binding protein (MBP), a hepatic lectin specific for mannose and N-acetylglucosamine, were investigated by utilizing gene recombinant technology. The major findings obtained in this sutdy are as follows.
1. The primary structure of rat liver MBP was deduced from its cDNA sequence. cDNA clones encoding rat liver MBP were isolated from a rat liver cDNA library carried in gt 11 by screening with an affinity purified anti-rat liver MBP antibody. The nucleotide sequence of the cDNA determined by the dideoxymethod revealed the complete amino acid sequence of the MBP (226 residues). The NH_2-terminal residue of the MBP, glutamic acid, was preceded by a signal peptide consisting of hydrophobic strecth of 18 amino acids. Near the NH_2-terminal, there was a collagen-like domain, which consisted of 19 repeats of the sequence, Giy-X-Y. The COOH-terminal half of the molecule obtained by collagenase digestion retained the sugar binding activity, revealing that the carbohydrate recognizing domain is located in its COOH terminal 150 amino acid residues. Treatment with reducing agents inactivated the lectin but this effect was reversed by air oxidation, indicating that disulfide bonds are associated with the expression of the binding activity.
2. Identification of endogenous liginds of the liver MBP. Endogenous ligands for the hepatic lectin were isolated from rat liver rough microsomes and primary cultured hepatocytes by affinity chromatography on an immobilized MBP column. Western blotting using specific antisera revealed that biosynthetic intermediates of serum glycoproteins, alpha_1-acid glycoprotein, alpha_1-macroglobulin, and alpha_1-antitrypsin, and a lysosomal enzyme,beta-glucuronidase were the major constituents of the endogenous ligands. These results suggest that the liver MBP is associated with the transport of glycoproteins within the cells during the process of their biosynthesis.

  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Kawasaki,T.: Carbohydrate Research. 151. 197-206 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Oka,S.: Journal of Biochemistry. 101. 135-144 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ikeda,K.: Journal of Biological Chemistry. 262. 7451-7454 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Oka,S.: Archives of Biochemistry and Biophysics. 260. 257-266 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mori,K.: Archives of Biochemistry and Biophysics. 264. 647-656 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ii,M.: Biochemical and Biophysical Research Communication. 155. 720-725 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ii,M.: Journal of Biochemistry. 104. 730-734 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawasaki,T.: Methods in Enzymology.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawasaki,T.: Methods in Enzymology.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawasaki,T.: "Vertebrate Lectins" Van Nostrand Reinhold, 10 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 川嵜敏祐: "レセプターの化学" 化学同人, 11 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 川嵜敏祐: "細胞工学 特集 動物レクチンとその機能" 秀潤社, 8 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 川嵜敏祐: "実験医学 特集 蛋白質の細胞内輸送とリーティング" 羊土社, 5 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kawasaki, T.: "Isolation and characterization of a receptor lectin specific for galactose/N-acetylgalactosamine from macrophages." Carbohydrate Research. 151. 197-206 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Oka, S.: "Primary structure of rat liver mannan-binding protein deduced from its cDNA sequence." Journal of Biochemistry. 101. 135-144 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ikeda, K.: "Serum lectin with known structure activates complement through the classical pathway." Journal of Biological Chemistry. 262. 7451-7454 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Oka,S.: "Isolation and characterization of two distinct mannan-binding proteins from rat serum." Archives of Biochemistry and Biophysics. 260. 257-266 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mori, K.: "Isolation and characterization of endogenous ligands for liver mannan-binding protein." Archives of Biochemistry and Biophysics. 265. 647-656 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ii, M.: "Structural similarity between the macrophage lectin specific for galactose/N-acetylgalactosamine and the hepatic asialo-glycoprotein binding protein." Biochemical and Biophysical Research Communication. 155. 720-725 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ii, M.: "Isolation and characterization of lectins specific for mannose/fucose/N-acetylglucosamine from peritoneal macrophages." Journal of Biochemistry. 104. 730-734 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawasaki, T.: "Isolation of mannose/N-acetylglucosamine binding protein proteins from mammalian sera." Methods in Enzymology. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawasaki, T.: "Isolation of a galactose/N-acetylgalactosamine binding protein and mannose/l-fucose/N-acetylglucosamine binding proteins from rat peritoneal macrophages." Methods in Enzymology. in press.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawasaki, T.: Vertebrate Lectins. Van Nostrand Reinhold, 10 (1987)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1990-03-20  

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