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2003 Fiscal Year Final Research Report Summary

Molecular analysis on activity regulation of growth factors by a new glycosaminoglycan.

Research Project

Project/Area Number 14572029
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionCHIBA UNIVERSITY

Principal Investigator

TOIDA Toshihiko  Chiba University, Grad.Sch.Pharmceu.Sci., Professor, 大学院・薬学研究院, 教授 (60163945)

Project Period (FY) 2002 – 2003
KeywordsAcharan sulfate / Lewis lung carcinoma cells / C57BL / 6 mice / Ceruloplasmin / Nucleolin
Research Abstract

Acharan sulfate (AS) is a new type of glycosaminoglycan from the giant African snail, Achatina fulica. AS having a primarily repeating disaccharide structure of α-D-Nacetylglucosaminyl 2-sulfoiduronate was studied as an antitumor agent using both in vivo and In vitro assays. We have found that this glycosaminoglycan binds specifically to serum ceruloplasmin. We also investigated the antitumor activity of AS in vivo as well as In vivo. To confirm the localization of exogeneous AS after s.c. injection in tumor of C57BL/6 mice implanted with Lewis lung carcinoma cells (LLC), the sectioned tissue was stained by Alcian blue/periodic acid-Schiff reagent. Exogenous AS was found to be localized mainly on the outer membrane of tissues. In vitro binding assay indicated that the binding of AS to. cells was markedly increased when 50 μg/ml of the sample was incubated for 5 hr. In the immunofluorescence assay using anti-AS antibody, AS was also detected around the cell surface membrane. Based on the fact that AS can bind the protein on cancer cell surface, the outer cell surface was biotinylated to identify any paricular binding protein. The biotinylated cells were lysed and the lysates were fractionated on AS affinity column with a stepwise salt gradient. Each fraction was visualized by SDS-PAGE and Western blotting. We focused on the protein eluted at 1 M NaCl and detected only two bands of which the molecular weights are approximately 118,000 and 52,000 Da. By ESI Q-TOF mass spectrometry, one band representing 110 kDa was determined as nucleolin, which is in a phosphorylated form on the cell surface of different cells as a cell surface receptor for various ligands, especially growth factors such as bFGF and midkines. The above results suggest that AS could show the inhibitory effects against tumor growth by. binding to specific receptors of cell surface.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Toida T, et al.: "Purification and characterization of heparan sulfate peptidoglycan from bovine liver."Carbohydr.Polym.. 48. 153-160 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Pretreatment procedure for the microdetermination of CS in plasma and urine."Anal.Biochem.. 302. 169-174 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Effect of(1-->3)- and (1-->4)-linkages of fully sulfated polysaccharides on their anticoagulant activity."Carbohydr.Res.. 337. 925-933 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Purification and characterization of dermatan sulfate from the skin of the eel, Anguilla"Carbohydr.Res.. 338. 263-269 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Chondroitin O-methyl ester : an unusual substrate for chondroitin AC lyase."Carbohydr.Res.. 338. 2101-2104 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Effects of Low Molecular Weight Chondroitin Sulfate on Type II Collagen-Induced Arthritis in DBA/1J Mice."Biol.Pharm.Bull.. 27. 47-51 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toida T, et al.: "Purification and characterization of heparan sulfate peptidoglycan from bovine liver."Carbogydr.Polym.. 48. 153-160 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toida T., et al.: "Pretreatment procedure for the microdetermination of chondroitin sulfate in plasma and urine."Anal.Biochem.. 302. 169-174 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toida T., et al.: "Effect of (1-->3)-and (1-->4)-linkages of fully sulfated polysaccharides on their anticoagulant activity."Carbohydr Res.. 337. 925-933 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toida T., et al.: "Purification and characterization of dermatan sulfate from the skin of the eel, Anguilla japonica."Carbohydr.Res.. 338. 263-269 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toida T., et al.: "Chondroitin O-methyl ester : an unusual substrate for chondroitin AC lyase."Carbohydr.Res.. 338(20). 2101-2104 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toida T., et al.: "Effects of Low Molecular Weight Chondroitin Sulfate on Type II Collagen-Induced Arthritis in DBA/1J Mice."Biol.Pharm.Bull.. 27. 47-51 (2004)

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

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Published: 2005-04-19  

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