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Solid-State NMR Study of Structure-Function Relationship of Protein Induced in the Signal Transduction Pathway at the Membrane Surface

Research Project

Project/Area Number 15570164
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionUniversity of Hyogo (2004)
Himeji Institute of Technology (2003)

Principal Investigator

TUZI Satoru  University of Hyogo, Graduate School of Life Science, Department of Life Science, Associate Professor, 大学院・生命理学研究科, 助教授 (60227387)

Co-Investigator(Kenkyū-buntansha) YAMAGUCHI Satoru  University of Hyogo, Graduate School of Life Science, Department of Life Science, Research Associate, 大学院・生命理学研究科, 助手 (20347529)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywordssolid state NMR / phospholipase C / PH domain / signal transduction pathway / lipid bilayer / biomembrane / peripheral membrane protein / higher order structure
Research Abstract

In 2003, we have established a technique of introduction of ^<13>C isotope labels into PLC-δ1 PH domain, and performed structural study of the PH domain at the membrane surface by using the solid state ^<13>C NMR spectroscopy. In 2004, (1)changes of the membrane binding structure of the PH domain induced at the negatively charged membrane surface were investigated, and (2)methods of ^<13>C isotope labeling of the remaining domains (EF-hand, XY, and C2) of PLC-δ1 were established.
The structure of the PH domain was found to be strongly influenced by acidic lipid contents of the membranes. In the presence of 20% of acidic lipid, phosphatidylserine, PH domain takes a structure similar to that in the solution instead of the previously reported membrane-binding structure observed at the electrically neutral membrane surface. The structural changes observed by the solid-state ^<13>C NMR spectroscopy revealed that the electrostatic repulsions between the negatively charged membrane surface and the charged residues of the PH domain prevent non-specific hydrophobic interaction between the amphipathic α2-helix of the PH domain and the hydrophobic inner layer of the membrane which causes the unique membrane-binding structure of the PH domain at the neutral membrane surface. Increase in the mobility of the PH domain was also observed at the negatively charged membrane surface. Those dependences of the dynamic structure of the PH domain on the lipid composition of membrane might related to responses of the PLC-δ1 to physiological changes of local lipid compositions such as a formation of lipid micro domain and a change of asymmetry of lipid bilayer.
Methods of ^<13>C labeling and preparation of solid state NMR samples of the PH-EF fragment, EF-hand domain and intact PLC-δ1 have been established in this year. These samples will be utilized to investigate inter-domain interactions between PLC-δ1 domains at the membrane surface.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (8 results)

All 2005 2003 Other

All Journal Article (4 results) Book (3 results) Publications (1 results)

  • [Journal Article] Structure and Dynamics of the Phospholipase C-δ1 Pleckstrin Homology Domain Located at the Lipid Bilayer Surface2003

    • Author(s)
      Satoru Tuzi
    • Journal Title

      The Journal of Biological Chemistry 278

      Pages: 28019-28025

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] イノシトールリン脂質結合タンパク質の脂質認識機構と膜結合による構造変化2003

    • Author(s)
      八木澤 仁
    • Journal Title

      細胞工学 22

      Pages: 859-864

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Structure and Dynamics of the Phospholipase C-δ1 Pleckstrin Homology Domain Located at the lipid Bilayer Surface2003

    • Author(s)
      Satoru Tuzi
    • Journal Title

      J.Biol.Chem. 278

      Pages: 28019-28025

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phosphoinositide-binding Proteins : To What Extent Do They See Their Substrates or Ligands, and How Dynamic Their Structures Change Upon Interaction with Membranes?2003

    • Author(s)
      Hitoshi Yagisawa
    • Journal Title

      Cell Tech. 22

      Pages: 859-864

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] Handbook of Modern Magnetic Resonance2005

    • Author(s)
      Satoru Tuzi
    • Publisher
      Kluwer Academic Publishers
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] Handbook of Modem Magnetic Resonance2005

    • Author(s)
      Satoru Tuzi
    • Publisher
      Kluwer Academic Publishers
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] The Handbook of Modern Magnetic Resonance2005

    • Author(s)
      Satoru Tuzi, Naoko Uekama, Masashi Okada, Hitoshi Yagisawa(共著)
    • Publisher
      Kluwer Academic Publishers
    • Related Report
      2004 Annual Research Report
  • [Publications] Satoru Tuzi: "Structure and Dynamics of the Phospholipase C-δ1 Pleckstrin Homology Domain Located at the Lipid Bilayer Surface"The Journal of Biological Chemistry. 278・30. 28019-28025 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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