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Reconstruction of signal transduction machinary in the cell membrane

Research Project

Project/Area Number 15KT0087
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeMulti-year Fund
Section特設分野
Research Field Constructive Systems Biology
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Sako Yasushi  国立研究開発法人理化学研究所, 開拓研究本部, 主任研究員 (20215700)

Project Period (FY) 2015-07-10 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000)
Fiscal Year 2017: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2016: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2015: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Keywordsシグナル伝達 / ナノマシン
Outline of Final Research Achievements

We have developed reconstitution membrane of EGF receptor (EGFR) for analyzing molecular mechanism of signal transduction across the plasma membrane. EGFR molecules were incorporated into nanodiscs with the diameter of 10-15 nm. Single-molecule FRET imaging of the transmembrane (TM)-juxtamenbrane (JM) domains of EGFR revealed that dimerization of EGFR, which is responsible for signal transduction, is induced by interactions of the JM domain with acidic membrane lipids, and inhibitory effect of the phosphorylation at the JM domain for dimerization requires acidic lipids in the membrane. Cholesterol in the membrane also induced dimerization of EGFR, however, dimer structures suggested to be different from that in membranes without cholesterol.

Academic Significance and Societal Importance of the Research Achievements

細胞膜受容体は細胞内外の情報変換を行う重要な蛋白質分子であるが、膜を介した情報伝達は信号分子と受容体の相互作用、受容体の分子構造・会合状態変化、受容体と細胞質分子の相互作用などが、さらに複雑かつ不均一な膜構造・膜脂質組成と絡み合って起こる複雑な反応である。ナノディスクは反応要素を最も小数・単純化できる再構成系であり、リポソームなど再構成膜系を利用した構成的システム研究へ基盤情報を与えることができる。本研究では蛋白質と脂質の協調が反応医御に必要であることが明らかになった。

Report

(5 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • 2015 Research-status Report
  • Research Products

    (35 results)

All 2019 2018 2017 2016 2015 Other

All Int'l Joint Research (4 results) Journal Article (13 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 13 results,  Open Access: 12 results,  Acknowledgement Compliant: 2 results) Presentation (15 results) (of which Int'l Joint Research: 4 results,  Invited: 15 results) Remarks (2 results) Patent(Industrial Property Rights) (1 results)

  • [Int'l Joint Research] Ulsan University(韓国)

    • Related Report
      2017 Research-status Report
  • [Int'l Joint Research] Incerm(France)

    • Related Report
      2016 Research-status Report
  • [Int'l Joint Research] McGill University(Canada)

    • Related Report
      2016 Research-status Report
  • [Int'l Joint Research] National University of Singapore(Singapore)

    • Related Report
      2016 Research-status Report
  • [Journal Article] Shift in Conformational Equilibrium Induces Constitutive Activity of G-Protein-Coupled Receptor, Rhodopsin.2018

    • Author(s)
      Maeda R, Hiroshima M, Yamashita T, Wada A, Sako Y, Shichida Y, Imamoto Y.
    • Journal Title

      J. Phys. Chem. B

      Volume: 122 Issue: 18 Pages: 4838-4843

    • DOI

      10.1021/acs.jpcb.8b02819

    • NAID

      120006708207

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Automated single-molecule imaging in living cells2018

    • Author(s)
      Yasui Masato、Hiroshima Michio、Kozuka Jun、Sako Yasushi、Ueda Masahiro
    • Journal Title

      Nature Communications

      Volume: 9 Issue: 1 Pages: 1-11

    • DOI

      10.1038/s41467-018-05524-7

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule diffusion-based estimation of ligand effects on G protein?coupled receptors2018

    • Author(s)
      Yanagawa Masataka、Hiroshima Michio、Togashi Yuichi、Abe Mitsuhiro、Yamashita Takahiro、Shichida Yoshinori、Murata Masayuki、Ueda Masahiro、Sako Yasushi
    • Journal Title

      Science Signaling

      Volume: 11 Issue: 548 Pages: 1-16

    • DOI

      10.1126/scisignal.aao1917

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-Molecule F?rster Resonance Energy Transfer Measurement Reveals the Dynamic Partially Ordered Structure of the Epidermal Growth Factor Receptor C-Tail Domain2018

    • Author(s)
      Okamoto Kenji、Sako Yasushi
    • Journal Title

      The Journal of Physical Chemistry B

      Volume: 123 Issue: 3 Pages: 571-581

    • DOI

      10.1021/acs.jpcb.8b10066

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule imaging and manipulation of biomolecular machines and systems2018

    • Author(s)
      Iino Ryota、Iida Tatsuya、Nakamura Akihiko、Saita Ei-ichiro、You Huijuan、Sako Yasushi
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - General Subjects

      Volume: 1862 Issue: 2 Pages: 241-252

    • DOI

      10.1016/j.bbagen.2017.08.008

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Inferring a nonlinear biochemical network model from a heterogeneous single-cell time course data2018

    • Author(s)
      Shindo Yuki、Kondo Yohei、Sako Yasushi
    • Journal Title

      Scientific Reports

      Volume: 8 Issue: 1 Pages: 6790-6790

    • DOI

      10.1038/s41598-018-25064-w

    • NAID

      120006502573

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Transient Acceleration of Epidermal Growth Factor Receptor Dynamics Produces Higher-Order Signaling Clusters2018

    • Author(s)
      Hiroshima Michio、Pack Chan-gi、Kaizu Kazunari、Takahashi Koichi、Ueda Masahiro、Sako Yasushi
    • Journal Title

      Journal of Molecular Biology

      Volume: 430 Issue: 9 Pages: 1386-1401

    • DOI

      10.1016/j.jmb.2018.02.018

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Lipid-Protein Interplay in Dimerization of Juxtamembrane Domains of Epidermal Growth Factor Receptor2018

    • Author(s)
      Maeda Ryo、Sato Takeshi、Okamoto Kenji、Yanagawa Masataka、Sako Yasushi
    • Journal Title

      Biophysical Journal

      Volume: 114 Issue: 4 Pages: 893-903

    • DOI

      10.1016/j.bpj.2017.12.029

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Single-molecule fluorescence-based analysis of protein conformation, interaction, and oligomerization in cellular systems2017

    • Author(s)
      Okamoto Kenji、Hiroshima Michio、Sako Yasushi
    • Journal Title

      Biophysical Reviews

      Volume: 10 Issue: 2 Pages: 317-326

    • DOI

      10.1007/s12551-017-0366-3

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Recent advances in FRET for the study of protein interactions and dynamics.2017

    • Author(s)
      Okamoto, K. and Sako, Y.
    • Journal Title

      Curr. Opin. Strct. Biol.

      Volume: 46 Pages: 16-23

    • DOI

      10.1016/j.sbi.2017.03.010

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] A novel spingomyeling/cholesterol specific probe reveals the dynamics of the membrane domain during virus and in Nieman-Pick type C2016

    • Author(s)
      Makino, A., Sako, Y. et al.
    • Journal Title

      FASEB J.

      Volume: 31 Issue: 4 Pages: 1301-1322

    • DOI

      10.1096/fj.201500075r

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cortical polarity of the RING protein PAR-2 is maintained by exchange rate kinetics at the cortical-cytoplasm boundary2016

    • Author(s)
      Arata, Y., Hiroshima, M., Pack, C.-G., Ramanujam, R., Motegi, F., Nakazato, K., Wiseman, P. W., Sawa, H., Kobayashi, T. J., Brandao, H. B., Shibata, T., and Sako, Y.
    • Journal Title

      Cell Rep.

      Volume: 16 Issue: 8 Pages: 2156-2168

    • DOI

      10.1016/j.celrep.2016.07.047

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] State transition analysis of spontaneous branch migration of the Holliday junction by photon-based single-molecule fluorescence resonance energy transfer.2016

    • Author(s)
      Okamoto, K. and Sako, Y.
    • Journal Title

      Biophys. Chem.

      Volume: 209 Pages: 21-27

    • DOI

      10.1016/j.bpc.2015.11.004

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Sinlgle-molecular pharmacology of GPCRs.2019

    • Author(s)
      Sako, Y.
    • Organizer
      The 3rd Biosignal Research Center International Symposium “Modulation of GPCR Signaling by Membrane Heterogeneity and Molecular Clustering”
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] In cell automated single-molecule analysis and its extensive applications.2018

    • Author(s)
      Hiroshima, M., Yasui, M., Kozuka, J., Sako, Y., and Ueda, M.
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞膜受容体の運動・会合と生理活性2018

    • Author(s)
      佐甲靖志
    • Organizer
      第17回生化学セミナー、東邦大学医学部
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] . Single-molecule pharmacology of a GPCR2017

    • Author(s)
      Yanagawa, M., Hiroshima, M., Togashi, Y., Yamashita, T., Shichida, Y., Murata, M., Ueda, M., and Sako, Y.
    • Organizer
      University of Strasbourg-RIKEN Workshop on Membrane Lipidology
    • Place of Presentation
      Strasbourg (France)
    • Year and Date
      2017-03-10
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 単一分子計測で明らかにする細胞中の分子動態2017

    • Author(s)
      岡本憲二、廣島通夫、日比野佳代、佐甲靖志
    • Organizer
      レーザー学会学術講演会第37回年次大会
    • Place of Presentation
      徳島大学(徳島)
    • Year and Date
      2017-01-07
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] 細胞膜受容体の1分子動態、生理学から薬理学へ2017

    • Author(s)
      佐甲靖志
    • Organizer
      日本顕微鏡学会 第42回関東支部講演会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Lipid-protein cooperativity in the regulation of juxtamembrane domain dimer formation in epidermal growth factor receptor2017

    • Author(s)
      Maeda, R., Sato, T., Okamoto, K., and Sako, Y.
    • Organizer
      第55回日本生物物理学会年会
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] Single-molecule FRET measurement for EGFR-RAS-MAPK signal transduction pathway.2017

    • Author(s)
      Okamoto, K., Maeda, R., and Sako, Y.
    • Organizer
      Deciphering complex energy landscape and kinetic network from single molecules to cells: a new challenge to make theories meet experiments
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research / Invited
  • [Presentation] 細胞内情報伝達のイメージング2017

    • Author(s)
      佐甲靖志
    • Organizer
      理研公開シンポジウム「観る・測る・解く」4次元細胞計測の現状と未来
    • Related Report
      2017 Research-status Report
    • Invited
  • [Presentation] 細胞内の1分子イメージング:生理学から薬理学へ2016

    • Author(s)
      佐甲靖志
    • Organizer
      東京理科大学研究推進機構総合研究院 イメージングフロンティアセンターシンポジウム
    • Place of Presentation
      東京理科大学(野田)
    • Year and Date
      2016-12-10
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] 細胞膜受容体の1分子可視化2016

    • Author(s)
      佐甲靖志
    • Organizer
      第25回日本バイオイメージング学会学術集会
    • Place of Presentation
      名古屋市立大学(名古屋)
    • Year and Date
      2016-09-05
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] 細胞内の確率過程2016

    • Author(s)
      佐甲靖志
    • Organizer
      研究会「異常拡散をめぐる最近の進展」
    • Place of Presentation
      慶應義塾大学(横浜)
    • Year and Date
      2016-09-01
    • Related Report
      2016 Research-status Report
    • Invited
  • [Presentation] 1分子計測で見るEGF受容体の細胞膜動態2016

    • Author(s)
      佐甲靖志
    • Organizer
      蛋白研セミナー
    • Place of Presentation
      大阪大学、吹田
    • Year and Date
      2016-03-04
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] 細胞膜受容体の動的会合体形成と機能2015

    • Author(s)
      佐甲靖志
    • Organizer
      第13回JCGGシンポジウム
    • Place of Presentation
      ウインク愛知、名古屋
    • Year and Date
      2015-10-01
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] Single-molecule analysis of cell signaling reactions2015

    • Author(s)
      佐甲靖志
    • Organizer
      第3回 少数性成物学トレーニングコース
    • Place of Presentation
      大阪大学、吹田
    • Year and Date
      2015-08-01
    • Related Report
      2015 Research-status Report
    • Invited
  • [Remarks] http://www.riken.go.jp/cell-info/

    • Related Report
      2016 Research-status Report
  • [Remarks] 佐甲細胞情報研究室Webpage

    • URL

      http://www.riken.go.jp/cell-info/

    • Related Report
      2015 Research-status Report
  • [Patent(Industrial Property Rights)] Gタンパク質共役型受容体(GPCR)の活性評価方法2017

    • Inventor(s)
      柳川正隆、佐甲靖志、廣島通夫、安井真人、上田昌宏、冨樫祐一
    • Industrial Property Rights Holder
      柳川正隆、佐甲靖志、廣島通夫、安井真人、上田昌宏、冨樫祐一
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2017-084803
    • Filing Date
      2017
    • Related Report
      2017 Research-status Report

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Published: 2015-07-14   Modified: 2020-03-30  

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