Elucidating the function of GPCR dimerization by single-molecule observation

• Project/Area Number: 24770120
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Functional biochemistry
• Research Institution: Kyoto University
• Principal Investigator: KASAI Rinshi 京都大学, 再生医科学研究所, 助教 (20447949)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
• Keywords: 生物物理 / 1分子計測 / Gタンパク質共役型受容体 / ダイマー形成 / １分子計測

Research Abstract

As a result of the single fluorescent molecule observation of beta 2 adrenergic receptor (b2AR), we found that 1) it's under the dynamic dimer-monomer equilibrium in the plasma membrane even at the resting state, 2) the dimer lifetime is ~80 milliseconds, 3) agonist stimulation increases the dimer lifetime by 40%. By observing trimeric G-protein together with b2AR at the single molecule level, it was further found that trimeric G-protein temporarily binds to b2AR monomers as well as dimers with comparable lifetimes, ~20 milliseconds. These reversible binding-unbinding processes were not affected by agonist stimulation. The results obtained here indicate that G-protein signaling is regulated in such a dynamic manner.

Research Products

• [Journal Article] Single-molecule imaging revealed dynamic GPCR dimerization (2014)
  • Author(s): Kasai RS、Kusumi A
  • Journal Title: Current Opinion in Cell Biology Volume: Vol. 27 Pages: 78-86
  • Peer Reviewed
• [Journal Article] Single-molecule imaging revealed dynamic GPCR dimerization (2014)
  • Author(s): R. S. Kasai and A. Kusumi
  • Journal Title: Curr Opin Cell Biol. Volume: 27 Pages: 78-86
  • DOI: 10.1016/j.ceb.2013.11.008
  • Peer Reviewed
• [Journal Article] ABCA1 dimer-monomer interconversion during HDL generation revealed by single-molecule imaging (2013)
  • Author(s): Nagata KO、Nakada C、Kasai RS、Kusumi A、Ueda K
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: Vol. 110 Pages: 5034-5039
  • Peer Reviewed
• [Journal Article] Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging (2013)
  • Author(s): Nishimura H、Ritchie K、Kasai RS、Goto M、Morone N、Sugimura H、Tanaka K、Sase I、Yoshimura A、Nakano Y、Fujiwara TK、Kusumi A
  • Journal Title: Journal of Cell Biology Volume: Vol. 202 Pages: 967-983
  • Peer Reviewed
• [Journal Article] Single-Molecule Imaging of Receptor–Receptor Interactions (2013)
  • Author(s): Suzuki KGN、Kasai RS、Fujiwara TK、Kusumi A
  • Journal Title: Methods in Cell Biology Volume: Vol. 117 Pages: 373-390
  • Peer Reviewed
• [Journal Article] Biocompatible fluorescent silicon nanocrystals for single-molecule tracking and fluorescence imaging (2013)
  • Author(s): Nishimura H, Ritchie K, Kasai RS, Morone N, Sugimura H, Tanaka K, Sase I, Yoshimura A, Nakano Y, Fujiwara TK, and Kusumi A
  • Journal Title: J Cell Biol. Volume: 202 Issue: 6 Pages: 967-83
  • DOI: 10.1083/jcb.201301053
  • Peer Reviewed
• [Journal Article] Single-molecule imaging of receptor-receptor interactions (2013)
  • Author(s): K. G. Suzuki, R. S. Kasai, T. K. Fujiwara, and A. Kusumi
  • Journal Title: Methods Cell Biol. Volume: 117 Pages: 373-390
  • DOI: 10.1016/b978-0-12-408143-7.00020-7
  • ISBN: 9780124081437
  • Peer Reviewed
• [Journal Article] ABCA1 dimer-monomer interconversion during HDL generation revealed by single-molecule imaging. (2013)
  • Author(s): Nagata, K., O., Nakada, C.,, Kasai, R. S.,Kusumi, A. and Ueda, K.
  • Journal Title: Proc Natl Acad Sci USA Volume: 110 Issue: 13 Pages: 5034-5039
  • DOI: 10.1073/pnas.1220703110
  • Peer Reviewed
• [Journal Article] Raf inhibitors target ras spatiotemporal dynamics (2012)
  • Author(s): Cho KJ、Kasai RS、Park JH、Chigurupati S、Heidorn SJ、van der Hoeven D、Plowman SJ、Kusumi A、Marais R、Hancock JF
  • Journal Title: Current Biology Volume: Vol. 22 Pages: 945-955
  • Peer Reviewed
• [Journal Article] Dynamic organizing principles of the plasma membrane that regulate signal transduction : commemorating the fortieth anniversary of Singer and Nicolson's fluid-mosaic model (2012)
  • Author(s): Kusumi A、Fujiwara TK、Chadda R、Xie M、Tsunoyama TA、Kalay Z、Kasai RS、Suzuki KGN
  • Journal Title: Annual Review of Cell and Developmental Biology Volume: Vol. 28 Pages: 215-250
  • Peer Reviewed
• [Journal Article] Membrane mechanisms for signal transduction : the coupling of the meso-scale raft domains to membrane-skeleton-induced compartments and dynamic protein complexes (2012)
  • Author(s): Kusumi A、Fujiwara TK、Morone N、Yoshida KJ、Chadda R、Xie M、Kasai RS、Suzuki KGN
  • Journal Title: Seminars in Cell & Developmental Biology Volume: Vol. 23 Pages: 164-144
  • Peer Reviewed
• [Journal Article] Transient GPI-anchored protein homodimers are units for raft organization and function (2012)
  • Author(s): Suzuki KGN、Kasai RS、Hirosawa KM、Nemoto YL、Ishibashi M、Miwa Y、Fujiwara TK、Kusumi A
  • Journal Title: Nature Chemical Biology Volume: Vol. 8 Pages: 774-783
  • Peer Reviewed
• [Journal Article] Transient GPI-anchored protein homodimers are units for raft organization and function (2012)
  • Author(s): K. G. N. Suzuki, R. S. Kasai, K. M. Hirosawa, Y. L. Nemoto, M. Ishibashi, Y. Miwa, T. K. Fujiwara, and A. Kusumi
  • Journal Title: Nat. Chem. Biol. Volume: 8 Issue: 9 Pages: 774-783
  • DOI: 10.1038/nchembio.1028
  • Peer Reviewed
• [Journal Article] Raf inhibitors target Ras spatiotemporal dynamics (2012)
  • Author(s): K. J. Cho, R. S. Kasai, J. H. Park, S. Chigurupati, S. J. Heidorn, D. van der Hoeven, S. J. Plowman, A. Kusumi, R. Marais, and J. F. Hancock
  • Journal Title: Curr. Biol. Volume: 22 Issue: 11 Pages: 945-955
  • DOI: 10.1016/j.cub.2012.03.067
  • Peer Reviewed
• [Presentation] GPCR dimers as active signal triggers : inverse agonist effects revealed by single-molecule imaging analysis (2013)
  • Author(s): Kasai RS, Fujiwara TK, Kusumi A
  • Organizer: 第51回日本生物物理学会年回
  • Place of Presentation: 京都
  • Year and Date: 2013-10-30
• [Presentation] Transient GPCR dimers trigger signaling as revealed by single-molecule imaging (2013)
  • Author(s): Kasai RS, Kusumi A
  • Organizer: The 2013 Cold Spring Harbor Asia Conference"NEW ADVANCES IN OPTICAL IMAGING OF LIVE CELLS & ORGANISMS"
  • Place of Presentation: 蘇州中国
  • Year and Date: 2013-08-22
• [Presentation] Dynamic Monomer-Dimer Equilibrium as a General Property of the Class A GPCR : Single-Molecule Imaging Study (2013)
  • Author(s): Kasai RS
  • Organizer: The 14th Membrane Research Forum
  • Place of Presentation: 京都
  • Year and Date: 2013-03-16
• [Presentation] Transient GPCR dimers trigger signaling as revealed by single-molecule imaging (2013)
  • Author(s): Kasai RS, Kusumi A
  • Organizer: The 2013 Cold Spring Harbor Asia Conference “NEW ADVANCES IN OPTICAL IMAGING OF LIVE CELLS & ORGANISMS”
  • Place of Presentation: 蘇州 中国
  • Invited
• [Presentation] GPCR dimers as active signal triggers: inverse agonist effects revealed by single-molecule imaging analysis (2013)
  • Author(s): Rinshi S. Kasai, Takahiro K. Fujiwara, and Akihiro Kusumi
  • Organizer: 日本生物物理学会 第51回年回
  • Place of Presentation: 京都
• [Presentation] Dynamic Monomer-Dimer Equilibrium as a General Property of the Class A GPCR: Single-Molecule Imaging Study (2013)
  • Author(s): Kasai RS
  • Organizer: The 14th Membrane Research Forum
  • Place of Presentation: 京都
  • Invited
• [Presentation] Dynamic monomer-dimer equilibrium as a general property of the class A GPCR : detection by single-molecule imaging (2012)
  • Author(s): Kasai RS, Fujiwara TK, Kusumi A
  • Organizer: The 2012 American Society for Cell Biology Annual Meeting / ASCB 52th Annual Meeting
  • Place of Presentation: サンフランシスコアメリカ
  • Year and Date: 2012-12-16
• [Presentation] Single-molecule imaging revealed acceleration of trimeric G-protein turnover by transient dimerization of ligandedβ2-AR (2012)
  • Author(s): Kasai RS
  • Organizer: 第50回日本生物物理学会年回
  • Place of Presentation: 名古屋
  • Year and Date: 2012-09-22
• [Presentation] Dynamic monomer-dimer equilibrium as a general property of the class A GPCR: detection by single-molecule imaging (2012)
  • Author(s): Kasai RS, Fujiwara TK, Kusumi A
  • Organizer: The 2012 American Society for Cell Biology Annual Meeting
  • Place of Presentation: San Francisco, U.S.A.
• [Book] "1分子イメジングにより明らかになった, GPCRのモノマー・ダイマーの動的な交換"、Clinical Neuroscience月刊臨床神経科学 (2013)
  • Author(s): 笠井倫志
  • Publisher: 中外医学社
• [Book] 超高速・超解像1蛍光分子顕微鏡"、現代化学 (2013)
  • Author(s): 藤原敬宏、笠井倫志、楠見明弘
  • Publisher: 東京化学同人
• [Book] Annu. Rev. Cell Dev. Biol., Volume 28 (2012)
  • Author(s): Kusumi A, Fujiwara TK, Chadda R, Xie M, Tsunoyama TA, Kalay Z, Kasai RS, Suzuki KGN
  • Total Pages: 36
  • Publisher: Annual Reviews
• [Book] Semin. Cell Dev. Biol., Volume 23 (2012)
  • Author(s): Kusumi A, Fujiwara TK, Morone N, Yoshida KJ, Chadda R, Xie M, Kasai RS, Suzuki KGN
  • Total Pages: 19
  • Publisher: Elsevier
• [Remarks]
  • URL: http://www.nanobio.frontier.kyoto-u.ac.jp/