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Molecular Biological Study on the Macrophage Lectin

Research Project

Project/Area Number 04454593
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KAWASAKI Toshisuke  Dept.Biol.Chem., Fac.Pharm.Sci., Kyoto.U.Professor, 薬学部, 教授 (50025706)

Co-Investigator(Kenkyū-buntansha) KOZUTSUMI Y  Dept.Biol.Dhem., Fac.Pharm.Sci., Kyoto.U.Associate Professor, 薬学部, 助教授 (70205425)
伊藤 信行  京都大学, 薬学部, 教授 (10110610)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1992: ¥3,600,000 (Direct Cost: ¥3,600,000)
Keywordsmacrophage / lectin / endocytosis / internalization signal / phosphotyrosine / cDNA / recombinant lectin / galactose / ゲノム遺伝子 / エキソン / イントロン / チロシンリン酸化 / ホスファターゼ / ガラクトース / 細胞内移行シグナル / チロシン / 肝レクチン
Research Abstract

In our previous study, we found that the Gal/GalNAc-specific lectin on rat peritoneal macrophages (macrophage asialoglycoprotein binding protein, M-ASGP-BP) is structurally similar to rat hepatic lectin (RHL) and is highly homologous with the major component of RHL,RHL-1. In this study, we demonstrated that transfection with a cDNA clone which encodes a single polypeptide, M-ASGP-BP,was sufficient for the expression of an endocytotic receptor for asialoorosomucoid (ASOR) on the COS-1 cell surface. The Kuptake value for ASOR was 12.5 nM,which is similar to that of peritoneal macrophages (23 nM) , and the number of ASOR bound on the cell surface was about 5 x 10 5/cell, this being hundreds of times larger than that for peritoneal macrophages. Gel filtration study indicated that M-ASGP-BP is a hexamer or octamer of a single polypeptide chain of 42 kDa. The aminoterminus of M-ASGP-BP deduced from its cDNA sequence contained the sequence, Tyr5-Glu6-Asn7-Phe8, in its cytoplasmic tail. The role of this putative internalization signal in the cytoplasmic tail was studied by measuring the endocytic of the wild type and mutant M-ASGP-BPs expressed on COS-1 cells through transfection wiht the wild type and mutant cDNAs prepared by oligonucleotide-directed mutagenesis, respectively. On the deletion of Tyr5 of replacement of it with alanine, the internalization of ASOR decreased to approximately on fourth that in the case of wild type. These results suggest that the Tyr5 in the recombinant M-ASGP-BP is necessary for its rapid internalization. The gene organization of M-ASGP-BP was determined from the sequences of PCR amplified DNA fragments and genomic DNA clones. The rat M-ASGP-BP gene consists of ten exons separated by nine introns, and the overall exon-intron organization is haighly homologous with that of RHL-gene.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] Ozaki.K.,et al.: "Expression of a functional asialoglycoprotein receptor through transfection of a cloned cDNA that encodes a macrophage lectin." J.Biol.Chem.267. 9229-9235 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ozaki.K.,et al.: "Role of tyrosine-5 in the cytoplasmic tail of the macrophege asialoglycoprotein receptor in the rapid internalization of ligands." J.Biochem.113. 271-276 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Wada.M.,et al.: "Gene stucture of macrophage asialoglycoprotein-binding protein and its tissue distribution." 発表予定(J.Biochem.).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ozaki.K., Ii.M., Itoh.N.& Kawasaki.T.: "Expression of a functional asialoglycoprotein receptor through transfection of a cloned cDNA that encodes a macrophage lectin." J.Biol.Chem.267. 9229-9235 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ozaki.K., Itoh.N.& Kawasaki.T.: "Role of tyrosine-5 in the cytoplasmic tail of the macrophege asialoglycoprotein receptor in the rapid internalization of ligands." J.Biochem.113. 271-276 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Wada.M., et al.: "Gene structure of macrophage asialoglycoprotein-binding protein and its tissue distribution." J.Biochem.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ozaki,K.,et al.: "The difference in structural specificity for recognition and binding between asialoglycoprotein receptors of liver and macrophages." Glycoconjugate J.(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Keiihi Ozaki: "Role of tyrosine-5 in the cyto plasmic tail of the macrophage asialoglycoprotein receptor in the rapid internalization" J.Biochem. 113. 271-276 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Keiichi Ozaki: "Expression of a functional asialogly co protein receptor through transtection of a cloned cDNA that encodes a macrophage lectin." J.Biol.Chem.267. 9229-9235 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Keiichi Ozaki: "Role of tyrosine-5 in the cytoplasmic tail of the macrophage arialogly c o protein receptor in the rapid internalization of ligands." J.Biochem.113. (1993)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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