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Dynamic structural rearrangements of ion channels and receptors dependent on expression density.

Research Project

Project/Area Number 16390052
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionNational Institute for Physiological Sciences

Principal Investigator

KUBO Yoshihiro  National Institute for Physiological Sciences, Department of Molecular Physiology, Professor, 分子生理研究系, 教授 (80211887)

Co-Investigator(Kenkyū-buntansha) SATO Chikara  National Institute of Advanced Science and Technology, Neuroscience Research Institute, Group Leader, 脳神経情報研究部門, グループ長 (00357146)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥13,800,000 (Direct Cost: ¥13,800,000)
Fiscal Year 2005: ¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 2004: ¥7,400,000 (Direct Cost: ¥7,400,000)
KeywordsATP receptor / P2X / single particle structure analysis / phospholipids / structural rearrangement / FRET / structure-function relationship / functional ragulation / 膜電位依存性 / ゲート機構 / ATP受容体チャネル / P2X2 / バキュウロウイルス / Sf9細胞
Research Abstract

Our aim of research is to elucidate the mechanisms of the environment dependent changes of ion channels and receptor function and to know their dynamic structural rearrangements. During the period of this grant, we have conducted four specific researches in the following. [1]We, for the first time, observed that the pore properties and the ligand sensitivity of ATP receptor channel P2X2 change depending on "the open channel density on the membrane" probably by mutual interaction between them. We also uncovered by systematic mutagenesis study that Ile328 in the second transmembrane region is critical for the expression density dependent changes. [2]We purified recombinant protein of P2X2 by baculo virus - Sf9 cell expression system, and carried out stoichiometry analyses by anchoring the recombinant proteins with glutaraldehyde. We observed that P2X2 is composed of three subunits. We also performed single particle structure analysis using the recombinant proteins, and confirmed that P2X2 is a trimeric protein which shows an inverted pyramid like structure. [3]As a possible mechanism of the density dependent changes of P2X2, we analyzed phospholipids dependency. We observed that the P2X2 function depends not on PIP2 but on PIP3, and that the dilated pore state is directly linked to the desensitized state. [4]We analyzed the structural rearrangements of the metabotropic glutamate receptor by FRET technique, and observed that the relative allocation of the dimeric subunits changes upon glutamate application. We also observed that the type of coupling G proteins of mGluR1 changes depending on the type of ligand, showing that mGluR1 is not a simple on-/off- switch but a multiple path regulator.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (27 results)

All 2006 2005 2004

All Journal Article (27 results)

  • [Journal Article] Dual signaling is differentially activated by different active states of the metabotropic glutamate receptor1α.2006

    • Author(s)
      Tateyama, M., Kubo, Y.
    • Journal Title

      Proc. Natl. Acad. Sci. USA 103

      Pages: 1124-1128

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Functional roles of charged amino acid residues on the wall of the cytoplasmic pore of Kir2.1.2006

    • Author(s)
      Fujiwara, Y., Kubo, Y.
    • Journal Title

      Journal of General Physiology 127

      Pages: 401-419

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 代謝型グルタミン酸受容体の動的構造変化とシグナリングの多様性2006

    • Author(s)
      立山充博, 久保義弘
    • Journal Title

      細胞工学 25

      Pages: 247-250

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Dual signaling is differentially activated by different active states of the metabotropic glutamate receptorla.2006

    • Author(s)
      Tateyama, M., Kubo, Y.
    • Journal Title

      Proc.Natl.Acad.Sci.USA 103

      Pages: 1124-1128

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Dimeric rearrangement and multiple signaling of metaobotropic glutamate receptor.2006

    • Author(s)
      Tateyama, M., Kubo, Y.
    • Journal Title

      Cell Technology 25

      Pages: 247-250

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Dual signaling is differentially activated by different active states of the metabotropic glutamate receptor1a.2006

    • Author(s)
      Tateyama, M., Kubo, Y.
    • Journal Title

      Proc.Natl.Acad.Sci.USA 103

      Pages: 1124-1128

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Functional roles of charged amino acid residues on the wall of the cytoplasmic pore of Kir2.1.2006

    • Author(s)
      Fujiwara, Y., Kubo, Y.
    • Journal Title

      Journal of General Physiology (印刷中)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Visualization of the trimeric P2X_2 receptor with a crown-capped extracellular domain.2005

    • Author(s)
      Mio, K., Kubo, Y., Ogura, T., Yamamoto, T., Sato, C.
    • Journal Title

      Biochemical and Biophysical Research Communications 337

      Pages: 998-1005

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Protein Kinase C shifts the voltage-dependence of KCNQ/M channels expressed in Xenopus oocytes.2005

    • Author(s)
      Nakajo, K., Kubo, Y.
    • Journal Title

      Journal of Physiology 569

      Pages: 59-74

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Towards a view of functioning dimeric metabotropic receptors2005

    • Author(s)
      Kubo, Y., Tateyama, M.
    • Journal Title

      Current Opinion in Neurobiology 15

      Pages: 289-295

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] International Union of Pharmacology. LIV. Nomenclature and molecular relationships of Kir channels.2005

    • Author(s)
      Kubo, Y. 他9名.
    • Journal Title

      Pharmacological Reviews 57

      Pages: 509-526

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 内向き整流性K^+ チャネルIRK1の構造機能連関とその遺伝子異常による疾患2005

    • Author(s)
      久保義弘, 藤原祐一郎
    • Journal Title

      別冊 医学のあゆみ 「イオンチャネル最前線 update」(倉智嘉久編) Suppl.215

      Pages: 88-93

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 代謝型グルタミン酸受容体の構造と機能調節機構2005

    • Author(s)
      久保義弘, 立山充博
    • Journal Title

      Clinical Neuroscience 24

      Pages: 149-152

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Towards a view of functioning dimeric metabotropic receptors.2005

    • Author(s)
      Kubo, Y., Tateyama, M.
    • Journal Title

      Current Opinion in Neurobiology 15

      Pages: 289-295

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] International Union of Pharmacology. LIV. Nomenclature and molecular relationships of Kir channels.2005

    • Author(s)
      Kubo, Y.et al.(10 authors in total)
    • Journal Title

      Pharmacological Reviews 57

      Pages: 509-526

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Structure-function relationship and channelopathy of inward rectifier potassium channel IRK1.2005

    • Author(s)
      Kubo, Y., Fujiwara, Y.
    • Journal Title

      Igaku-no-Ayumi Suppl. 215

      Pages: 88-93

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Structure and functional regulation of the metabotropic glutamate receptor2005

    • Author(s)
      Kubo, Y, Tateyama, M.
    • Journal Title

      Clinical Neuroscience 24

      Pages: 149-152

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Towards a view of functioning dimeric metabotropic receptors2005

    • Author(s)
      Kubo, Y, Tateyama, M.
    • Journal Title

      Current Opinion in Neurobiology 57

      Pages: 509-526

    • Related Report
      2005 Annual Research Report
  • [Journal Article] OPA1 expression in the normal rat retina and optic nerve.2005

    • Author(s)
      Ju, W.K., Misaka, T., Kushnareva, Y., Nakagomi, S, Agarwal, N., Kubo, Y.et al.
    • Journal Title

      Journal of Comparative Neurology 488

      Pages: 1-10

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Visualization of the trimeric P2×2 receptor with a crown-capped extracellular domain.2005

    • Author(s)
      Mio, K., Kubo, Y., Ogura, T., Yamamoto, T., Sato, C.
    • Journal Title

      Biochemical and Biophysical Research Communications 337

      Pages: 998-1005

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Density dependent changes of the pore properties of P2X_2 receptor channel.2004

    • Author(s)
      Fujiwara, Y., Kubo, Y.
    • Journal Title

      Journal of Physiology 558

      Pages: 31-43

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ligand-induced rearrangement of the intracellular dimeric conformation of metabotropic glutamate receptor1α.2004

    • Author(s)
      Tateyama, M., Abe, H., Nakata, H., Saitoh, O, Kubo, Y.
    • Journal Title

      Nature Structural & Molecular Biology 11

      Pages: 637-642

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Density Dependent Changes of the Pore Properties of P2X_2 Receptor Channel.2004

    • Author(s)
      Fujiwara, Y, Kubo, Y.
    • Journal Title

      Journal of Physiology 558

      Pages: 31-43

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ligand-induced rearrangement of the intracellular dimeric conformation of metabotropic glutamate receptor1a.2004

    • Author(s)
      Tateyama, M., Abe, H., Nakata, H., Saitoh, O, Kubo, Y.
    • Journal Title

      Nature Structural & Molecular Biology 11

      Pages: 637-642

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ligand-induced rearrangement of the intracellular dimeric conformation of metabotropic glutamate receptorla.2004

    • Author(s)
      Tateyama, M., Abe, H., Nakata, H., Saitoh, O, Kubo, Y.
    • Journal Title

      Nature Structural & Molecular Biology 11

      Pages: 637-642

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Density Dependent Changes of the Pore Properties of P2X_2 Receptor Channel.2004

    • Author(s)
      Fujiwara, Y., Kubo, Y.
    • Journal Title

      Journal of Physiology 558

      Pages: 31-43

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Biochemical and Physiological Analyses of the RGS8 function2004

    • Author(s)
      Saitoh, O, Kubo, Y.
    • Journal Title

      Methods in Enzymology 390

      Pages: 129-148

    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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