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Elucidatiop of Tangle Formation Mechanism Based on 3D-structural Analysis of Tau Protein

Research Project

Project/Area Number 13680752
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionOsaka University of Pharmaceutical Sciences

Principal Investigator

ISHIDA Toshimasa  Osaka University of Pharmaceutical Sciences, Pharmacy, Professor, 薬学部, 教授 (00111021)

Co-Investigator(Kenkyū-buntansha) MINOURA Katsuhiko  Osaka University of Pharmaceutical Sciences, Pharmacy, Assistant, 薬学部, 助手 (10278591)
TOMOO Koji  Osaka University of Pharmaceutical Sciences, Pharmacy, Lecturer, 薬学部, 講師 (70257898)
IN Yasuko  Osaka University of Pharmaceutical Sciences, Pharmacy, Assistant, 薬学部, 助手 (50257896)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
Keywordstau protein / formation mechanism of tangle / microtubule-binding domain / 3D-structure / X-ray analysis / NMR / Spectroscopic analysis / molecular modeling / タングル形式
Research Abstract

1)X-ray crystal structure analysis of microtubule-binding domain
To elucidate 3D structure of tau protein, we attempted various crystallizations of tau and its fragments. Consequently, we succeeded in the crystallization of tau microtubule-binding domain (MBD) as GST fusion protein, which were prepared as recombinant by gene expression from E.coli. The crystal data are : tetragonal, space group P4_32_12, a=b=92.24Å, c=57.68Å. X-ray diffraction data up to 2Å resolution were collected by synchrotron of Spring 8. The crystal structure was solved by the molecular replacement method using the 3D structure of GST. After several structure refinement, the present R factor is 0.258. Although the MBDs formed dimeric structure, they showed large conformational flexibility and their atomic positions were not determined. This is in contrast with the rigid 3D structure of GST. Since the MBD constructs the core structure of tau PHF, the present result indicates the close relationship between the confo … More rmational flexibility of MBD and PHE formation. We are now in progress in analyzing the crystal structure more accurately.
2)Analyses of solution structure of MBD
MBD is composed of three-or four-repeated structure, where each repeat peptide (named R1 -R4) consists of 31 or 32 amino acid residues. To elucidate the conformational flexibility of each repeat peptide, the solution structures of R1 -R4 in TEE solution were analyzed by the combination of NMR and model building. Consequently, the the structures of R1, R2 and R4 peptides showed α-helical structure of amphipathic distribution of respective amino acid residues. In contrast, R3 took both of extended and α-helical structures of amphipathic behavior. This result suggest the molecular association of MBD through alternative hydrophilic and hydrophobic interactions.
3)Investigation of MBD interactions and model of repeat-structure-dependent PHF formation
The association of neighboring MBDs were investigated by ThS fluorescence, light scattering and surface plasmon resonance analyses, and electromicroscopy. On the basis of these results, we proposed the first model of repeat structure-controlled step-by-step filament formation of 4RMBD. This model would be helpful upon considering rational approaches to prevent tau deposition in AD Less

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] K.Minoura, T.Tomoo, T.Ishida, H.Hasegawa, M.sasaki, T.Taniguchi: "Amphipathic helical behaviour of the third repeat fragment in the tau microtubule-binding domain, studied by ^1H NMR spectroscopy."Biochem.Biophys.Res.Commun.. 294. 210-214 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, K.Tomoo, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Solvent-dependent conformation of the third repeat fragment in microtubule-binding domain of tau protein, analyzed by 1H-NMR spectroscopy and molecular modeling calculation"Bul.Chem.Soc.Jpn. 76. 1617-1624 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.-M.Yao, K.Tomoo, T.Ishida, M.Sumida, M.sasaki, T.Taniguchi: "Aggregation analysis of the microtubule binding domain in tau protein by spectroscopic method"Peptide Science. 2002. 249-252 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.In, H.Ohishi, T.Ishida, Y.Igarashi: "Concerted interaction between conjugated double bond CHs and multiple OHs in polyene macrolide antibiotic chainin : weak =C-...O interactions responsible for intrinsic molecular assembly"ChemCommun. 2003. 1692-1693 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.-M.Yao, K.Tomoo, T.Ishida, H.Hasegawa, M.sasaki, T.Tanibuchi: "Aggregation analysis of the microtubule binding domain in tau protein by spectroscopic methods"J.Biochem.. 134. 91-99 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, T.-M.Yao, K.Tomoo, M.Sumida, M.Sasaki, T.Taniguchi, T.Ishida: "Different associational and conformational behaviors between the second and third repeat fragments in the tau microtubule-binding domain"Eur.J.Biochem.. 271. 545-552 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hiraoka, T.-M.Yao, K.Minoura, K.Tomoo, M.Sumida, T.Taniguchi, T.Ishida: "Conformational transition state is responsible for assembly of microtubule-binding domain of tau protein"Biochem.Biophys.Res.Commun.. 315. 659-663 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, K.Tomoo, Y.In, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Amphipathic helical behavior of the third repeat fragment in the tau microtuble-binding domain, studied by 1H-NMR spectroscopy"Biochem.Biophys.Res.Commun.. 294. 210-214 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, K.Tomoo, Y.In, T.ishida, H.hasegawa, M.Sasaki, T.Taniguchi: "Conformational study of repeat fragments in microtuble binding domain of tau protein"Peptide Science. 2002. 329-332 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.-M.Yao, K.Tomoo, Y.In, T.Ishida, M.Sumida, M.sasaki, T.Taniguchi: "Aggregation analysis of microtubule binding domain in tau protein by spectroscopic method"Peptide Science. 2002. 249-252 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] T.-M.Yao, K.Tomoo, T.Ishida, H.Hasegawa, M.sasaki, T.Tanibuchi: "Aggregation analysis of the microtubule binding domain in tau protein by spectroscopic methods"J.Biochem.. 134. 91-99 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, K.Tomoo, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Solvent-dependent conformation of the third repeat fragment in the microtubule-binding domain of tau protein, analyzed by 1H-NMR spectroscopy and molecular modeling calculation"Bull.Chem.Soc.Jpn. 76. 1617-1624 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Y.In, H.Ohishi, T.Ishida, Y.Igarashi: "Concerted interaction between conjugated double bond CHs and multiple OHs in polyene macrolide antibiotic chainin : weak =C-H...O interactions responsible for intrinsic molecular assembly"Chem.Commun.. 2003. 1692-1693 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, T.-M.Yao, K.Tomoo, M.Sumida, M.Sasaki, T.Taniguchi, T.Ishida: "Different associational and conformational behaviors between the second and third repeat fragments in the tau microtubule-binding domain"Eur.J.Biochem.. 271. 545-552 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Tomoo, T.-M.Yao, S.Hiraoka, M.Tokimasa, M.Sumita, M.Sasaki, T.Taniguchi, T.Ishida: "Analysis of aggrebation mechanism of microtubule binding domain in tau protein"Peptide Science. 2003. 303-306 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, T.-M.Yao, K.Tomoo, M.Sumida, T.Taniguchi, T.Ishida: "Conformational analysis of four repeat fragments in tau microtubule binding domain"Peptide Science. 2002. 371-374 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] S.Hiraoka, T.-M.Yao, K.Minoura, K.Tomoo, M.Sumida, T.Taniguchi, T.Ishida: "Conformational transition state is responsible for assembly of microtubule-binding domain of tau protein"Biochem.Biophys.Res.Commun.. 315. 659-663 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Minoura, K.Tomoo, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Solvent-dependent conformation of the third repeat fragment in the microtubule-binding domain of tau protein, analyzed by 1H-NMR spectroscopy and molecular modeling calculation"Bull.Chem.Soc.Jpn. 76. 1617-1624 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Y.In, H.Ohishi, T.Ishida, Y.Igarashi: "Concerted interaction between conjugated double bond CHs and multiple OHs in polyene macrolide antibiotic chainin : weak =C-H...O interactions responsible for intrinsic molecular assembly"Chem Commun. 2003. 1692-1693 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.-M.Yao, K.Tomoo, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Aggregation analysis of the microtubule binding domain in tau protein by spectroscopic methods"J.Biochem.. 134. 91-99 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] T.-M.Yao, K.Tomoo, T.Ishida, M.Sumida, M.Sasaki, T.Taniguchi: "Aggregation analysis of microtubule binding domain in tau protein by spectroscopic methods"Peptide Science. 2002. 249-252 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Minoura, T.-M.Yao, K.Tomoo, M.Sumida, M.Sasaki, T.Taniguchi, T.Ishida: "Different associational and conformational behaviors between the second and third repeat fragments in the tau microtubule-binding domain"Eur.J.Biochem.. 271. 545-552 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] S.Hiraoka, T.-M.Yao, K.Minoura, K.Tomoo, M.Sumida, T.Taniguchi, T.Ishida: "Conformational transition state is responsible for assembly of microtubule-binding domain of tau protein"Biochem.Biophys.Res.Commun.. 315. 659-663 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Minoura, K.Tomoo, T.Ishida, H.Hasegawa, M.Sasaki, T.Taniguchi: "Amphipathic Helical Behavior of the third Repeat Fragment in the Tau Microtubule-Binding Domain, Studied by ^1H-NMR Spectroscopy"Biochem. Biophys. Res. Commun.. 294. 210-214 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.-M.Yao, Tomoo, Y.In, H.Hasegawa, M.Sasaki, T.Taniguchi, T.Ishida: "Aggregation Analysis of Microtubule Binding Domain in Tau Protein by Spectroscopic Method"J. Biochem.. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Minoura, K.Tomoo, H.Hasegawa, M.Sasaki, T.Taniguchi, T Ishida: "Solvent-dependent conformation of the third repeat fragment in microtubule-binding domain of tau protein, analyzed by ^1H-NMR spectroscopy and molecular modeling calculation"Bull. Chem. Soc. Jpn. 76(in press). (2003)

    • Related Report
      2002 Annual Research Report

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

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