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1995 Fiscal Year Final Research Report Summary

Biosynthesis and Complex Formation of Cartilage Specific Functional Matrix/Chondromodulin-I

Research Project

Project/Area Number 06454655
Research Category

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

Allocation TypeSingle-year Grants
Research Field Functional biochemistry
Research InstitutionOsaka University

Principal Investigator

SUZUKI Fujio  Osaka Univ.Fac.of Dentistry, Professor, 歯学部, 教授 (40028717)

Co-Investigator(Kenkyū-buntansha) HIRAKI Yuji  Osaka Univ.Fac.of Dentistry, Assoc.Prof., 歯学部, 助教授 (40144498)
Project Period (FY) 1994 – 1995
KeywordsChondrocyte / Growth Factor / Cartilage Matrix / Chondromodulin-I / Processig / Cell Differentiation
Research Abstract

The complete primary amino acid sequence of chondromodulin-I (ChM-I) has been known for bovine counterpart. In the present study, we tried to clone out human ChM-I cDNA from which the primary amino acid sequence would be deduced. While cartilage tissue is widely found in embryonic tissue, there is only a little cartilage tissue found in postnatal animals. Most of cartilage in embryo is replated by bone at the end of embryonic development. Thus, there is a considerable difficulty in obtaining fresh cartilage tissue of the amount enough for construction of cDNA library. To avoid this difficulty, we isolated RNA from human chondrosarcoma of a highly differentiated type from which cDNA library was constructed. The full coding region of human ChM-I cDNA was successfully amplified from the cDNA by PCR.Short5' untranslated stretch was cloned out from human genomic DNA library. Sequencing of the human ChM-I cDNA fragment indicated that human ChM-I has three base/one amino acid deletion in comp … More arison with the bovine counterpart. Sequence identity between human and bovine Chm-I precursor was determined to be 89.6% based on the nucleotide sequences and 91.9% based on the deduced amino acid sequences. N-Glycosylation site in the mature ChM-I was conserved in the human counterpart, but one of two O-glycosylated Thr residues in bovine ChM-I was missing. In contrast, C-terminal half of the mature ChM-I which contains 8 cysteine residues was completely identical except for the one amino acid substitution (His to Tyr). Then, we attempted to express human ChM-I cDNA in COS cells. The yield of the expressed recombinant ChM-I was unexpectedly low. However, the yield was improved by replacing the 5' franking sequences of the ATG start codon from GGCTTC to GGCACC.We could identify mature human ChM-I in the conditioned medium from the transfected COS cell culture as a diffuse 25 kDa band by SDS-PAGE analysis. These results suggested that we could successfully establish the experimental model to study biosynthetic and secretory pathway of mature ChM-I. Less

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Hiraki, Y.: "Molecular cloning of a new class of cartilage-specific matrix, chondromodulin-I, which stimulates growth of cultured chondrocytes." Biochem. Biophys. Res. Commun.175. 971-977 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Suzuki, F.: "Regulation of cartilage metabolism." Bone and Mineral Research. 8. 115-142 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiraki, Y.: "Bifunctional role of chondromodulin-I (ChM-I) in endochondral bone formation." Intermational Symposium on Cartilage Metabolism Osaka, 1994(Abstract). 37-39 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takigawa, M.: "Cartilage-derived anti-tumor factor (CATF): Partial purification and correlation of inhibitory activity against tumor growth with anti-angiogenic activity." J. Bone Miner. Metab.6. 83-92 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohba, Y.: "Purification of an angiogenesis inhibitor from culture medium conditioned by a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8." Biochim. Biophys. Acta. 1245. 1-8 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hiraki, Y.: "Bone morphogenetic proteins (BMP-2 and BMP-3) promote growth and expression of the differentiated phenotype of rabbit chondrocytes and osteoblastic MC3T3-E1 cells in vitro." J. Bone Miner. Res.6. 1373-1385 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 開 祐司: "骨形成と骨吸収、及びそれらの調節因子2巻" 廣川書店, 10 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Hiraki: "Molecular cloning of a new class of cartilage-specific matrix, chondrocmodulin-I,which stimulates growth of cultured chondrocytes." Biochm.Biophys.Res.Commun.175. 971-977 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] F.Suzuki: "Regulation of cartilage matabolism" Bone and Mineral Research. 8. 115-142 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Hiraki: "Bifunctional role of chondromodulin-I (ChM-I) in endochondral bone formation." Intl.Symp.on Cartilage Metab.Osaka. 37-39 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M.Takigawa: "Cartilage-derived anti-tumor factor (CATF) : Partial purification and correlation of inhibitory activity against tumor growth with anti-angiogenic activity" J.Bone Min.Metab.6. 83-92 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Ohba: "Purification of an angiogenesis inhibitor from culture medium conditioned by a human chondrosarcoma-derived chondrocytic cell line, HCS-2/8." Biochim.Biophys.Acta. 1245. 1-8 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.Hiraki: "Bone morphogenetic proteins (BMP-2 and BMP-3) promote growth and expression of the differentiated phenotype of rabbit chondrocytes and osteoblastic MC3T3-E1 cells in vitro." J.Bone Min.Metab.6. 1373-1385 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y.HIRAKI: Hirokawa Shoten. Bone formation and resoption and their regulators, vol.2, 387-396 (1995)

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

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Published: 1997-03-04  

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