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Expression and function of RGD-CAP conatining RGD-motif in cartilage

Research Project

Project/Area Number 09671894
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional basic dentistry
Research InstitutionGrant-in-Aid for Scientific Research (C)(2)

Principal Investigator

NOSHIRO Mitsuhide  Hiroshima Univ.School of Dentistry, Associate Prof., 歯学部, 助教授 (00144858)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥2,500,000 (Direct Cost: ¥2,500,000)
Keywordscollagen / integrin / cartelage matrix / cloning / chondrocyte culture / cell adhesion / RGD-CAP / 軟骨細胞 / 細胞外基質 / cDNAクローニング
Research Abstract

The following results were obtained to clarify the role of RGD-CAP as the mediator in the interaction of cells and matrix concerning the growth and development of chondrocytes.
(1) A cDNA clone for RGD-CAP (collagen-associated protein containing the RGD sequence) was isolated from chick embryo chondrocyte cDNA library. The deduced amino acid sequence (680 amino acid residues) contained four internal repeat domains and two highly conserved sequences (Hi and H2) in each repeat. The protein contains an integrin recognition site, the RGD motif in the fourth domain of four repetitive structures.
(2) We examined the effect of recombinant chick RGD-CAP on the adhesion of chondrocytes and fibroblasts using RGD-CAP-coated dishes. When rabbit chondrocytes or human fibroblasts were seeded on plastic dishes coated with RGD- CAP, cell adhesion was enhanced compared with control dishes.
(3) RGD-CAP produced a synergistic stimulation of cell adhesion in thepresence of type II or III collagen, and an additive stimulation in the presence of type I or V collagen.
(4) The effect of RGD-CAP on cell adhesion required divalent cations (Mg^<2+> or Mn^<2+>) , and was reduced by EDTA and by the GRGDS peptide.
(5) Monoclonal antibodies to the human integrin alpha1, alpha5, or beta1 subunit but not alpha2, alpha3, or beta2 decreased the RGD-CAP-induced cell adhesion. These findings suggest that RGD-CAP is a novel ligand for integrin alpha1beta1/alpha5beta1, and that a site other than the RGD sequence on RGD-CAP is also involved in cell adhesion.
(6) An RGD-CAP fragment containing two repeating domains was also capable of enhancing efficiently the adhesion of chondrocytes indicating the importance of the repetition of two domains.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (34 results)

All Other

All Publications (34 results)

  • [Publications] Hashimoto,K.et al.: "Characterization of a cartilage-derived 66-kDa protein(RGD-CAP/βig-h3) that binds to collagen" Biochim.Biophys.Acta.1355. 303-314 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Okimura,A.et al.: "Cartilage-Matrix Protein(CMP) Synthesis is Enhanced in Arthritic Cartilage." Arthritis and Reumatism. 40. 1029-1036 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shen,M.et al.: "Molecular Characterization of the Novel Basic Helix-Loop-Helix Protein DEC1 Expressed in Differentiated Human Embryo Chondrocytes" Biochem.Biophys.Res.Commun.236. 294-298 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Noshiro,M.et al.: "Structural And Evolutional Studies On Sterol 14-Demethylase P450(CYP51),The Most Conserved P450 Monooxygenase" J.Biochem.122. 1114-1121 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshida,Y.et al.: "Structural and Evolutionary Studies on Sterol 14-Demethylase p450(CYP51),The Most Conserved P450 Monooxygenase:II.Evolutionary Analysis of Protein and Gene Structures" J.Biochem.122. 1122-1128 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Koyano,Y.et al.: "Cloning and Characterization of CDEP,a Novel Putative Rho Guanine Nucleotide Exchange Factor Expressed in Differentiated Human Embryonic Chondrocytes" Biochem.Biophys.Res.Commun,. 214. 369-375 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nishimura,M.,et al.: "Role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of extracellular matrices and fluids" Biochim.Biophys.Acta,. 1380. 1-9 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawamoto,T.,et al.: "Structural and phylogenetic analyses of RGD-CAP/βig-h3,a fasciclin-like adhesion protein expressed in chick chondrocyte" Biochim.Biophys.Acta,. 1395. 288-292 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawashima-Ohya,Y.et al.: "Effects of Parathyroid Hormone(PTH) and PTH-Related Peptide on Expressions of Matrix Metalloproteinases-2,-3 and -9 in Growth Plate Chondrocyte Cultures" Endocrinol. 139. 2120-2127 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawamoto,T.: "Expression of membrane-bound transferrin-like protein p97 on the surface of chondrocytes." Eur.J.Biochem. 256. 503-509 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawashima-Ohya,Y.et al.: "Retinol-Binding Protein is Produced in Rabbit Chondrocytes and Responds to Parathyroid Hormone(PTH)/PTH-Related Peptide-cAMP Pathway." Endocrinol.140in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Makihira,S.et al.: "Enhancement of Cell Adhesion and Spreading by a Cartilage-Characteristic Noncollagenous Protein,Cartilage-matrix Protein(CMP/Matrilin-1),via Integrinalbl" J.Biol.Chem.274in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hashimoto, K.et al.: "Characterization of a cartilage-derived 66-kDa protein (RGD-CAP/betaig-h3) that binds to collagen" Biochim.Biophys.Acta. 1355. 303-314 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Okimura, A.et al.: "Cartilage-Matrix Protein (CMP) Synthesis is Enhanced in Arthritic Cartilage." Arthritis and Reumatism. 40. 1029-1036 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shen, M.et al.: "Molecular Characterization of the Novel Basic Helix-Loop-Helix Protein DEC1 Expressed in Differentiated Human Embryo Chondrocytes" Bichem.Biophys.Res.Commun.236. 294-298 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Noshiro, M.et al.: "Structural And Evolutional Studies On Sterol 14-Demethylase P450 (CYP51), The Most Conserved P450 Monooxygenase" J.Biochem.122. 1114-1121 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshida, Y.et al.: "Structural and Evolutionary Studies on Sterol 14-Demethylase p450 (CYP51), The Most Conserved P450 Monooxygenase : II.Evolutionary Analysis of Protein and Gene Structures" J.Biochem.122. 1122-1128 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Koyano, Y.et al.: "Cloning and Characterization of CDEP,a Novel Putative Rho Guanine Nucleotide Exchange Factor Expressed in Differentiated Human Embryonic Chondrocytes" Biochem.Biophys.Res.Commun. 214. 369-375 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nishimura, M.et al.: "Role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of extracellular matrices and fluids" Biochim.Biophys.Acta. 1380. 1-9 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawamoto, T., et al.: "Structural and phylogenetic analyzes of RGD-CAP/betaig-h3, a fasciclin-like adhesion protein expressed in chick chondrocyte" Biochim.Biophys.Acta. 1380. 1-9 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawashima-Ohya, Y.et al.: "Effects of Parathyroid Hormone (PTH) and PTH-RelatedPeptide on Expressions of Matrix Metalloproteinases-2, -3 and -9 in Growth Plate Chondrocyte Cultures" Endocrinol. 139. 2120-2127 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawamoto, T.: "Expression of membrane-bound transferrin-like protein p97 on the surface of chondrocytes." Eur.J.Biochem. 256. 503-509 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawashima-Ohya, Y.et al.: "Retinol-Binding Protein is Produced in Rabbit Chondrocytes and Responds to Parathyroid Hormone (PTH)/PTH-Related Peptide-cAMP Pathway." Endocrinol.140(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Makihira, S.et al.: "Enhancement of Cell Adhesion and Spreading by a Cartilage-Characteristic Noncollagenous Protein, Cartilage-matrix Protein (CMP/Matrilin-1), via Integrin alpha1beta1" J.Biol Chem.(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawashima-Ohya,Y.et al.: "Effects of Parathyroid Hormone (PTH) and PTH-Related Peptide on Expressions of Matrix Metalloproteinases-2,-3 and -9 in Growth Plate Chondrocyte Cultures" Endocrinol. 139. 2120-2127 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kawamoto,T.: "Expression of membrane-bound transferrin-like protein p97 on the surface of chondroeytes." Eur.J.Biochem. 256. 503-509 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Aoyama,Y.et al.: "CYP51-Like Gene of Mycobacterium tuberculosis Actually Encodes a P450 Similar to Eukaryotic CYP51." J.Biochem.124. 694-696 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shiba,H.et al.: "Differential Effects of Various Growth Factors and Cytokines on the Syntheses of DNA,Type1 Collagen,Laminin,Fibronectin,Osteonectin/Secreted Protein,Acidic and Rich in Cystein (SPARC) and Alkaline Phosphatase by Human Pulp Cells in Culture" Journal of Cellular Physiology,. 174. 194-205 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kawashima-Ohya,Y.et al.: "Retinol-Binding Protein is Produced in Rabbit Chondrocytes and Responds to Parathyroid Hormone (PTH)/PTH-Related Peptide-cAMP Pathway." Endocrinol.140(in press). (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Makihira,S.et al.: "Enhancement of Cell Adhesion and Spreading by a Cartilage-Characteristic Noncollagenous Protein,Cartilage-matrix Protein (CMP/Matrilin-1),via Integrin α1β1" J.Biol Chem.274(in press). (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] K.Hashimoto, M.Noshiro, Y.Kato, et al.: "Characterization of a cartilage-derived 66-kDa protein (RGD-CAP/βig-h3)that binds to collagen." Biochim.Biophys.Acta.1355. 303-314 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Okimura, M.Noshiro, Y.Kato, et al.: "Cartilage-Matrix Protein(CMP) Synthesis is Enhanced in Arthritic Cartilage." Arthritis and Reumatism. 40. 1029-1036 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Nishimura, M.Noshiro, Y.Kato, et al.: "Role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of extracellular matrices and fluids." Biochim.Biophys.Acta. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Kawamoto, M.Noshiro, Y.Kato, et al.: "Structural and phylogenetic analyses of RGD-CAP/βig-h3,a fasciclin-like adhesion protein expressed in chick chondrocytes." Biochim.Biophys.Acta. (in press). (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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