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Super molecular association of laminin-8 creating contact structure between capillary endothelial cells and adipocytes.

Research Project

Project/Area Number 13460039
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionNagoya University

Principal Investigator

KITAGAWA Yasuo  Nagoya Univ. Grad. Sch. Bioagric. Sci., Professor, 大学院・生命農学研究科, 教授 (50101168)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥16,600,000 (Direct Cost: ¥16,600,000)
Fiscal Year 2002: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 2001: ¥12,400,000 (Direct Cost: ¥12,400,000)
Keywordslaminin / endothelial cells / adipocyte / angiogenesis / adipogenesis / heparin / 血管内皮細胞 / 脂肪細胞 / プラスミトゲン / プラスミノーゲン活性化酵素 / プラスミン / LG領域 / 脂肪形成 / プラスミノーゲン
Research Abstract

Laminin α4 chain is a component of laminin-8 (α4β1γ1) synthesized by endothelial cells and adipocytes. In order to study the function of laminin-8 during angiogenesis and adipogenesis, we focused on the G domain of α4 chain and found that the subdomain LG4 has highest affinity to heparin among LG1-LG5 subdomain forming the G domain. In order to specify the functional amino acid for the heparin-binding, we produced a fusion protein of α4LG4 with glutathione S-transferase in E. coli and introduced site-directed mutagenesis at 19 basic amino acids (K, R, H). Mapping of the functional basic amino acids on a 3D structure model of α4LG4 showed the heparin-binding surface locating at opposite side of the calcium-binding site. We also found that addition of recombinant α4LG1-LG5 is inhibitory to the de novo adipogenesis induced by subcutaneous injection of Matrigel and basic fibroblast growth factor into mice. α4LG4-5 and α4LG1-LG3 showed stronger inhibition than α4LG1-LG5, suggesting that the G domain of α4 chain in laminin-8 may be cleaved after secretion from endothelial cells and preadipocytes at the boundary between LG3 and LG4 for its function of supporting the adipogenesis by the interaction between endothelial cells and preadipocytes.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Tajima R, Toriyama K, Sano R, Inou K, Oda H, Toni S. Kitagawa Y.: "Reversible heat-denaturation of Matrigel retains biological activity inducing differentiated phenotype of primary cultured hepatocytes and inducing de novo adipogenesis"Tissue Engineering. (印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okazaki I, Suzuki N, Nishi N, Utani A, Matsuura H, Shinkai H, Yamashita H, KitagawaY, Nomizu M.: "Identification of biologically active sequences in the laminin alpha 4 chain G domain"J Biol Chem. 277(40). 37070-37078 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 後藤彰, 北川泰雄: "ポストゲノム時代におけるRNA干渉法の役割"バイオサエンスとインダストリー. 60(5). 26-29 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Toriyama K, Kawaguchi N, Kitoh J, Tajima R, Inou K, Kitagawa Y, Toni S.: "Endogenous adipocyte precursor cells for regenerative soft-tissorcupine stimulates the posttranslational N-glycosylation of wingless in the endoplasmic reticulum"Tissue Engineering. 8(1). 157-165 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tanaka K, Kitagawa Y, Kadowaki T: "Drosophila segment polarity gene product porcupine stimulates the posttranslational N-glycosylation of wingless in the endoplasmic reticulum"J Biol Chem.. 277(15). 12816-12823 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Goto A, Aoki M, Ichihara S. Kitagawa Y.: "α-,β-or γ-chain-specific RNA interference of laminin assembly in Drosophila Kc167 cells"Biochem J.. 360(1). 167-172 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tajima R, Toriyama K, Sano R, Inou K, Oda H, Torii S, Kitagawa Y: "Reversible heat-denaturation of Matrigel retains biological activity inducing differentiated phenotype of primary cultured hepatocytes and inducing de novo adipogenesis"Tissue Engineering. In press. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okazaki I, Suzuki N, Nishi N, Utani A, Matsuura H, Shinkai H, Yamashita H, Kitagawa Y, Nomizu M: "Identification of biologically active sequences in the laminin α4 chain G domain"J.Biol.Chem.. 277(40). 37070-37078 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Toriyama K, Kawaguchi N, Kitoh J, Tajima R, Inou K, Kitagawa Y, Torii S: "Endogenous adipocyte precursor cells for regenerative soft-tissorcupine stimulates the posttranslational N-glycosylation of wingless in the endoplasmic reticulum"Tissue Engineering. 8(1). 157-165 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Goto A, Aoki M, Ichihara S, Kitagawa Y: "α-, β- or γ-chain-specific RNA interference of laminin assembly in Drosophila Kc167 cells"Biochem.J.. 360(Pt1). 167-172 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tajima R, Kawaguchi N, Horino Y, Takahashi Y, Toriyama K, Inou K, Torii S, Kitagawa Y: "Hypoxic enhancement of type IV collagen secretion accelerates adipose conversion of 3T3-L1 fibroblasts"Biochim.Biophys.Acta. 1540(3). 179-187 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Tajima R, Toriyama K, Sano R, Inou K, Oda H, Torii S, Kitagawa Y.: "Reversible heat-denaturation of Matrigel retains biological activity inducing differentiated phenotype of primary cultured hepatocytes and inducing de novo adipogenesis"Tissue Engineering. (印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okazaki I, Suzuki N, Nishi N, Utani A, Matsuura H, Shinkai H, Yamashita H, Kitagawa Y, Nomizu M.: "Identification of biologically active sequences in the laminin alpha 4 chain G domain"J Biol Chem. 277(40). 37070-37078 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 後藤彰, 北川泰雄: "ポストゲノム時代におけるRAN干渉法の役割"バイオサイエンスとインダストリー. 60(5). 26-29 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Toriyama K, Kawaguchi N, Kitoh J, Tajima R, Inou K, Kitagawa Y, Torii S.: "Endogenous adipocyte Precursor cells for regenerative soft-tissorcupine stimulates the posttranslational N-glycosylation of wingless in the endoplasmic reticulum"Tissue Engineering. 8(1). 157-165 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tanaka K, Kitagawa Y, Kadowaki T: "Drosophila segment polarity gene product porcupine stimulates the posttranslational N-glycosylation of wingless in the endoplasmic reticulum"J Biol Chem.. 277(15). 12816-12823 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Tajima R, Kawaguchi N, Horino Y, Takahashi Y, Toriyama K, Inou K, Torii S, Kitagawa Y.: "Hypoxic enhancement of type IV collagen secretion accelerates adipose conversion of 3T3-L1 fibroblasts"Biochimica et Biophysica Acta. 1540(3). 179-187 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Akira Goto, Takeshi Kumagai, Chino Kumagai, Junko Hirose, Hiroshi Narita, Hitoshi Mori, Tatsuhiko Kadowaki, Konrad Beck, Yasuo Kitagawa: "A Drosophila haemocyte-specific protein, hemolectin, similar to human von Willebrand factor"Biochemical Journal. 359(1). 99-108 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Akira Goto, Makoto Aoki, Shigeyuki Ichihara, Yasuo Kitagawa: "α-, β- or γ-chain-specific RNA interference of laminin assembly in Drosophila Kc167 cells"Biochemical Journal. 360(1). 167-172 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 北川泰雄, 山下泰恒: "末梢組織におけるラミニンα鎖Gドメインの役割"生化学. 73(10). 1227-1233 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kazuhiro Toriyama, Nobuko Kawaguchi, Junzo Kitoh, Rie Tajima, Kazuhiko Inou, Yasuo Kitagawa, Shuhei Torii: "Endogenous adipocyte precursor cells for regenerative soft-tissue engineering"Tissue Engineering. 8(1). 157-165 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Tanaka K, Kitagawa Y, Kadowaki T.: "Drosophila segment polarity gene product porcupine stimulates the posttranslational N-glycosylation of wingless in the Endoplasmic reticulum"Journal of Biological Chemistry. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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