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Studies on the Chemical Structure of Enamel Proteins and the Mechanisms of Calcification

Research Project

Project/Area Number 01440077
Research Category

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

Allocation TypeSingle-year Grants
Research Field Functional basic dentistry
Research InstitutionTokyo Medical and Dental University

Principal Investigator

SASAKI Satoshi  Tokyo, Med. and Dent. University Dept. of Biochemistry, Faculty of Dentistry Professor, 歯学部生化学, 教授 (80013803)

Co-Investigator(Kenkyū-buntansha) SUZUKI Michiko  Tokyo Med. and Dent. University Dept. of Biochemistry, Faculty of Dentistry Tech, 歯学部生化学, 技官 (80171255)
SHIMOKAWA Hitoyata  Tokyo Med. and Dent. University Dept. of Biochemistry, Faculty of Dentistry Asso, 歯学部生化学, 助教授 (80014257)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥19,700,000 (Direct Cost: ¥19,700,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1990: ¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1989: ¥11,900,000 (Direct Cost: ¥11,900,000)
KeywordsEnamel / Calcification / Protein / Calcium
Research Abstract

Dental enamel is a highly mineralized tissue consisting almost exclusively of apatite crystals with a small amount of organic matrix. However, protein concentrations as high as 25-30 % can be found at an early stage of enamel formation. The protein components decrease gradually during the maturation of enamel and are replaced by the apatite crystals. The proteins present at the early stage of enamel formation, amelogenin and enamelin are thought to play important roles in the process of enamel mineralization.
1) We screened human genomic DNA library in phage EMBL3 with a bovine full-length cDNA probe and succeeded in isolating one clone containing a insert which encodes the human amelogenin. From the base sequence of the DNA fragments subcloned into several vectors, the primary structure of amelogenin protein was deduced except leader sequence, 1st and 2nd amino acids. However, it was discovered that human mature amelogenin protein consists of 178 amino acids.
2) We isolated an amelogenin-digestible enzyme having an acidic pH optimum from developing bovine enamel. This result suggested presence of acidic conditions in the enamel matrix. We tried to measure pH in developing teeth with color pH indicators and discovered the presence of cyclical acidic and neutral zones in developing enamel. When stained with pH indicators, unerupted bovine enamel manifested several alternating stripes of two colors corresponding to acidic (pH 5.8-6.0) and neutral (pH 7.0) reactions. Those pH values of the enamel samples from respective zone were confirmed by measuring with a ph-meter. The pH difference was detected in depth. Neutral bands corresponded to red stripes of GBHA-staining and acidic bands to unstained white zones. Importance of alternating acidic and neutral zones in relation to the functions of ruffle-ended and smooth-ended ameloblasts was confirmed for degradation of amelogenin matrix by acidic and neutral proteases and for growth and maturation of hydroxyapatite crystals.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Y.Ogata,H.Shimokawa,S.Sasaki: "Calcium binding activity of bovine enamelins" Dentistry in Japan. 26. 51-55 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T.Inage,H.Shimokawa,Y.Teranishi,T.Iwase,Y.Toda,I.Moro: "Immunocytochemical demonstration of amelogenin and enamelins secreted by ameloblasts during secretory and maturation stages" Arch.Histol.Cytol.52(3). 213-229 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Takgi,H.Shimokawa,M.Suzuki,H.Nagai,S.Sasaki: "Immunohistochemical localization of α 2HS-glycoprotein in dentin" Calcif.Tissue Int.47(1). 40-45 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M.Harada,M.Mogi,B.Y.Hiraoka,K.Hukawawa,T.Takagi,S.Sasaki: "Determination of proline iminopeptidase activity in bovine dental germs by HPLC" J.Chromatography. 527. 158-162 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S.Sasaki,T.Takagi,M.Suzuki: "Cyclical changes in pH in bovine developing enamel as sequential bands" Archs.Oral Biol.36(3). 227-231 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Ibaraki,H.Shimokawa,S.Sasaki: "An analysis of the biochemical and biosynthetic properties of dentin phosphoprotein" Matrix. 11. 115-124 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S.Sasaki,T.Takagi,M.Suzuki: "Mechanisms and Phyogeny of Mineralization in Biological Systems (eds.S.Suga,H.Nakahara) "Amelogenin Degradation by an Enzyme having Acidic pH Optimum and the Presence of Acidic Zone in Developing Bovine Enamel"" Springer-Verlag, 3 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Ogata, H. Shimokawa, S. Sasaki: "Calcium binding activity of bovine enamelins" Dentistry in Japan. 26. 51-55 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] T. Inage, H. Shimokawa, Y. Teranishi, T. Iwase, Y. Toda, I. Moro: "Immunohistochemical demonstration of amelogenins and enamelins secreted by ameloblasts during secretory and maturation stages" Arch. Histol. Cytol.52(3). 213-229 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Takagi, H. Shimokawa, M. Suzuki H. Nagai, S. Sasaki: "Immunohistochemical localizations of alpha2HS-glycoprotein in dentin" Calcif. Tissue Int.47(1). 40-45 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M. Harada, M. Mogi, B. Y. Hiraoka, K. Hukasawa, T. Takagi, S. Sasaki: "Determination of proline iminopeptidase activity in bovine dental germs by HPLC" J. Chromatography. 527. 158-162 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Sasaki, T. Takagi, M. Suzuki: "Cyclical changes in pH in bovine Developing enamel as sequential bands" Archs. Oral Biol.36(3). 227-231 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K. Ibaraki, H. Shimokawa, S. Sasaki: "An analysis of the biochemical and biosynthetic properties of dentin phosphoprotein" Matrix. 11. 115-124 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Takagi,H.Shimokawa,M.Suzuki H.Nagai,S.Sasaki: "Immunohistochemical localization of α2HSーglycoprotein in dentin" Calcif.Tissue Int.47(1). 40-45 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Harada,M.Mogi,B.Y.Hiraoka,K.Hukawawa,T.Takagi,S.Sasaki: "Determination of proline iminopeptidase activity in bovine dental germs by HPLC" J.Chromatography. 527. 158-162 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Sasaki,T.Takagi,M.Suzuki: "Cyclical changes in pH in bovine developing enamel as sequential bands" Archs.Oral Biol.36(3). 227-231 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Ibaraki,H.Shimokawa,S.Sasaki: "An analysis of the biochemical and biosynthetic properties of dentin phosphoprotein" Matrix. 11. 115-124 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Sasaki,T.Takagi,M.Suzuki: "Mechanisms and Phylogeny of Mineralization in Biological Systems(eds.S.Suga,H.Nakahara) “Amelogenin Degradation by an Enzyme having Acidic pH Optimum and the Presence of Acidic Zone in Developing Bovine Enamel"" SpringerーVerlag, 3 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Goseki,S.Oida,A.Nifuji,S.Sasaki: "Properties of Alkaline Phosphatase of the Human Dental Pulp" J.Dental Research. 69(3). 909-912 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] M.Goseki,S.Oida,Y.Tokagi,T.Okuyama,S.Sasaki: "Immunological Study of Hypophosphatasia" Clinica Chimica Acta. 190. 263-268 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 鈴木 ミチ子、佐々木 哲: "骨の基質に関する最近の知見" 腎と骨代謝. 3(4). 423-428 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 五関 正江、大井田 新一郎、佐々木 哲: "骨の加水分解酵素" 日本臨床. 48(12). 2828-2832 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Shimokawa,M.Tamura,K.Ibaraki,S.Sasaki: "Tooth Enamel V Human amelogenin gene" Florence Publishers, 545 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 須賀 昭一編,佐々木 哲: "図説齲蝕学ー歯の化学形成ー" 医歯薬出版, 310 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 下川仁弥太、荊木京未、田村正人、佐々木哲: "オステオネクチンの発現に関する研究" 日本骨代謝学会雑誌. 7(1). 21-25 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 佐々木哲: "骨の基質をめぐって" 日本歯科医師会雑誌. 42(5). 513-519 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 佐々木哲: "骨基質の生化学ー序論" BONE. 3(3). 29-31 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Y.Ogata,H.Shimokawa,S.Sasaki: "Calcium Binding Activity of Bovine Enamelin" Dentistry in Japan. 26. 51-55 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Y.Takagi,H.Shimokawa,M.Suzuki,H.Nagai,S.Sasaki: "Proceeding of the Third International Conference on the Chemistry and Biology of Mineralized Tissues,Immunohistochemical localization of alpha 2HSーGlycoprotein" Gordon and Breach Science Publishers, 334 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] H.Shimokawa,M.Tamura,K.Ibaraki,S.Sasaki: "Tooth Enamel V Human amelogenin gene" Florence Publishers, 545 (1989)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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