• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Analysis of molecular mechanism of hypophosphatasia

Research Project

Project/Area Number 14571759
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional basic dentistry
Research InstitutionNIIGATA UNIVERSITY

Principal Investigator

ODA Kimimitsu  NIIGATA UNIVERSITY, Graduate School of Medical and Dental Sciences, Professor, 大学院・医歯学総合研究科, 教授 (10122681)

Co-Investigator(Kenkyū-buntansha) ITO Masahiro  NIIGATA UNIVERSITY, Graduate School of Medical and Dental Sciences, Assistant, 大学院・医歯学総合研究科, 助手 (40313522)
AMAYA Yoshihiro  NIIGATA UNIVERSITY, Graduate School of Medical and Dental Sciences, Associate Professor, 大学院・医歯学総合研究科, 助教授 (50193032)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsHypophosphatasia / Alkaline phosphatase / Defect of Calcification / Calcium / polyubiquitination / proteasome / inborn error of bone metabolism / 遺伝子疾患 / 石灰化 / 先天性代謝異常 / ユビキチン / 分解
Research Abstract

Two missense mutations(N153D,D289V) of tissue-nonspecific alkaline phosphatase(TNSALP) gene associated with perinatal hypophosphatasia were analyzed. First off, specific mutations were introduced into the wild-type TNSALP cDNA using site-directed mutation and resultant plasmids were transfected into COS-1 cells. The cells expressing each TNSALP mutant was examined biochemically and morphologically.
1.Contrast to dimeric structure of the wild type, each TNSALP mutant was found to form a disulfide-bonded high-molecular-mass aggregate, indicative of defective folding and incorrect assembly. Resultantly, each TNSALP mutant exhibited no measurable alkaline phosphatase acitivity.
2.Both TNSALP mutants failed to reach the cell surface. Instead, TNSALP(D289V) was found to be localized mainly to the endoplasmic reticulum, while TNSALP(N153D) was in the cis-Golgi.
3.Both TNSALP mutants were degraded in the proteasome at different rates, presumably reflecting difference in their intracellular localization.
4.TNSALP(D289V) was found to be polyubiqutinated prior to degradation in the proteasome.
5.Aspartic acid at position 289 of TNSALP is assumed to be involved in the binding of calcium ion. So we analyzed another TNSALP mutant(E218G), which is also involved in coordination of calcium ion TNSALP mutant(E218G) quite resembled TNSALP(D289V) regarding to their molecular properties. These results raise the possibility that calcium binding is critical for acquisition of three dimensional structure of TNSALP.
Taken together, sever forms of hypophosphatasia caused by two missense mutations(N153D,D289V) of TNSALP can be classified as folding disease.

Report

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

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Ishida Y., et al.: "Tissue-nonspecific alkaline phosphatase with an Asp289-Val mutation fails to reach the cell surface and undergoes proteasome-mediated degradation"Journal of Biochemistry. 134-1. 65-70 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Amizuka N., et al.: "Defective bone remodeling in osteoprotegerin deficient mice"Journal of Electron Microscopy. 52-6. 503-513 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nakano Y., et al.: "Alkaline and acid phosphatases in bone cells serve as phosphohydrolases at physiological pH in vivo : histochemical evaluation"Connective Tissue Research. 44(suppl). 219-222 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hao H., et al.: "A histological evaluation on self-setting・-tricalcium phosphate applied in the rat bone cavity"Biomaterials. 25-3. 431-442 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ito M., et al.: "Retention at the cis-Golgi and delayed degradation of tissue-nonspecific alkaline phosphatase with an Asn153-Asp substitution, a cause of perinatal hypophosphatasia"Biochemical Journal. 361-3. 473-480 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kawai M., et al.: "Ectopic bone formation by human bone morphologenic protein-2 gene transfer to skeletal muscle using trancutaneous electroporation"Human Gene Therapy. 14-16. 1547-1556 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ishida Y., et al.: "Tissue-nonspecific alkaline phosphatase with an Asp289-Val mutation fails to reach the cell membrane and undergoes proteasome-mediated degradation"Journal of Biochmistry. 134-1. 65-70 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Amizuka N., et al.: "Defective bone remodeling in Osteoproteogerin deficient mice"Journal of Electron Microscopy. 52-6. 503-513 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Nakano Y., et al.: "Alkaline and acid phosphatase in bone cells serve as phosphohydrolases at physiological pH in vivo : histochemical evaluation"Connective Tissue Research. 44(Suppl). 216-222 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Hao H., et al.: "A histological evaluation on self-setting a-tricalcium phosphate applied in the rat bone cavity"Biomaterilas. 25-3. 431-442 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ito M., et al.: "Retention at the cis-Golgi and delayed degradation of tissue-non-specific alkaline phosphatase with an Asn153-Asp substitution, a cause of perinatal hypophosphatasia"Biochemical Journal. 361-3. 473-480 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kawai M., et al.: "Ectopic bone formation by human bone Morphologenic protein-2 gene transfer to skeletal muscle using Transcutaneous electroporation"Human Gene Therapy. 14-16. 1547-1556 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Ishida Y., et al.: "Tissue-nonspecific alkaline phosphatase with an Asp289-Val mutation fails to reach the cell surface and undergoes proteasome-mediated"Journal of Biochemistry. 134・1. 65-70 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Amizuka, N. et al.: "Defective bone remodeling in osteoprotegerin deficient mice"Journal of Electron Microscopy. 52・6. 503-513 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nakano Y., et al.: "Alkaline and acid phosphatases in bone cells serve as phosphohyrolases at physiological pH in vivo : histochemical evaluation"Connective Tissue Research. 44 suupl.. 219-222 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hao H., et al.: "A histological evaluation on self-setting α-tricalcium phosphate applied in the rat bone cavity"Biomaterials. 25・3. 431-442 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ito, M., et al.: "Jaw bone remodeling at the invasion front of gingival squamous cell carcinoma"Journal of Oral Pathology and Medicine. 32・1. 10-17 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kawai, M., et al.: "Ectopic bone formation by human bone morphologenic protein-2 gene Transfer to skeletal muscle using transcutaneous electroporation"Human Gene Therapy. 14・16. 1547-1556 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Amizuka, N.et al.: "Biological action of parathyroid hormone(PTH)-related peptide(PTHrP) mediated either by the PTH/PTHrP receptor or the nucleolar translocation"Anat. Sci. Int.. 77. 225-236 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ito, M.et al.: "Jaw bone remodeling at the invention front of gingival squamous cell carcinomas"J. Oral Path. Med.. 32. 10-17 (2003)

    • Related Report
      2002 Annual Research Report

URL: 

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

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi