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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
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥2,700,000 (Direct Cost: ¥2,700,000)
KeywordsAcharan sulfate / Lewis lung carcinoma cells / C57BL / 6 mice / Ceruloplasmin / Nucleolin / Lewis 肺ガン細胞 / 抗腫瘍活性 / 増殖因子 / 免疫系 / 表面プラズモン共鳴
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.

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [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
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Toida T., et al.: "Purification and characterization of heparan sulfate peptidoglycan from bovine liver."Carbohydr.Polym.. 48. 153-160 (2002)

    • Related Report
      2003 Annual Research Report
  • [Publications] Toida T., et al.: "Pretreatment procedure for the microdetermination of CS in plasma and urine."Anal.Biochem.. 302. 169-174 (2002)

    • Related Report
      2003 Annual Research Report
  • [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)

    • Related Report
      2003 Annual Research Report
  • [Publications] Toida T., et al.: "Purification and characterization of dermatan sulfate from the skin of the eel, Anguilla"Carbohydr.Res.. 338. 263-269 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Toida T., et al.: "Chondroitin O-methyl ester : an unusual substrate for chondroitin AC lyase."Carbohydr.Res.. 338. 2101-2104 (2003)

    • Related Report
      2003 Annual Research Report
  • [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)

    • Related Report
      2003 Annual Research Report
  • [Publications] Linhardt R.J., Toida T.: "Methods in Molecular Biology"Humana Press. 14 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Sakai S, Akiyama H, Toyoda H, Toida T, Maitani T, Imanari T.: "Effect of chondroitin sulfate on murine splenocytes sensitized with ovalbumin"Immunol Lett. 84. 211-216 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Islam T, Butler M, Sikkander SA, Toida T, Linhardt RJ.: "Further evidence that periodate cleavage of heparin occurs primarily through the antithrombin binding site"Carbohydr Res. 337. 2239-2243 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Linhardt RJ, Toida T.: "Tech.Sight. Capillary electrophoresis. Ultra-high resolution separation comes of age"Science. 298(5597). 1441-1442 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Yu G, Linhardt RJ, Toida T, Shadid H, Ferro V, Palermo MC, Podger D.: "Preparation and anticoagulant activity of the phosphosulfomannan PI-88"Eur J Med Chem. 37. 383-391 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ye S, Luo Y, Lu W, Linhardt RJ, Toida T, Pelletier H, McKeehan WL.: "Structural basis for interaction of FGF-1, FGF-2, and FGF-7 with different heparan sulfate motifs"Biochemistry. 40(48). 14429-14439 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai S, Toyoda H, Toida T, Imanari T, Linhardt RJ: "Pretreatment procedure for the microdetermination of chondroitin sulfate in plasma and urine"Anal Biochem. 302(2). 169-174 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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