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単一シリア内における動態と力学応答の共役を画像化する光学顕微鏡の開発

Publicly Offered Research

Project AreaCilium-centrosome system regulating biosignal flows
Project/Area Number 15H01218
Research Category

Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionGakushuin University

Principal Investigator

西坂 崇之  学習院大学, 理学部, 教授 (40359112)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥10,920,000 (Direct Cost: ¥8,400,000、Indirect Cost: ¥2,520,000)
Fiscal Year 2016: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2015: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Keywords分子モーター / 単一シリア / 光ピンセット / シグナル伝達 / 非対称運動 / 協調運動 / 単一シリアマニピュレーション
Outline of Annual Research Achievements

真核生物が発達させた特徴的な細胞器官である「シリア」は、メカニカルに溶媒の流れを生み出す装置でありながら、情報のフローをも制御するという驚くべき機能を持つ。シリア自体が情報を検知する例(不動繊毛など)もありながら、運動によって生じる流れが形態学的な特徴(ノードの動繊毛など)に直結するのだ、本課題では、これら躍動的なシリアの動態を精密に捉えるべく、単一シリアの内部における機能解析を可能にする新しい測定システムの構築を目指した。3次元の力学応答を精密に測定する独自技術をベースにし、最終的には外部からの摂動に対する機能応答の詳細を画像化する方法論を提案する。
平成28年度は以下の2つの測定に成功し、大きな研究の進展を見せた。① シリアの局所的な蛍光強度変化を測定する系の構築。励起レーザーを試料に対して急な角度で照射する遮光照明の技術により、背景光を極限まで抑え、蛍光蛋白からのシグナルを捉えることに成功した。従来の共焦点光学系を用いた観察方法に比べ、カメラの性能だけに依存した時間分解能を達成できるため汎用性が高く、また光ピンセットの光学系とも併用が可能である。② 3次元精密測定技術によるべん毛回転の高速撮影。繊毛の可視化に開発したシステムをべん毛による回転運動に応用し、微生物ハロバクテリウム・サリナラムのトルクの見積もりに成功した。直径 200 nm から 3μm までの蛍光ビーズをべん毛に直接結合し、菌体をガラス面に吸着させる。べん毛モーターの回転は、2次元では振動もしくは楕円軌道として観察されるが、再構成した3次元運動はきれいな円運動となった。正確な回転数を見積もることにより、アーキアモーターのトルクの精密測定に初めて成功した。

Research Progress Status

28年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

28年度が最終年度であるため、記入しない。

Report

(2 results)
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (19 results)

All 2017 2016 2015 2014

All Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 6 results,  Acknowledgement Compliant: 5 results) Presentation (10 results) (of which Int'l Joint Research: 6 results,  Invited: 7 results) Book (1 results)

  • [Journal Article] Dissection of the angle of single fluorophore attached to the nucleotide in corkscrewing microtubules2017

    • Author(s)
      Fujimura, S. Ito, Y. Ikeguchi, M. Adachi, K. Yajima, J. Nishizaka, T.
    • Journal Title

      Biochem Biophys Res Commun

      Volume: 485 Issue: 3 Pages: 614-620

    • DOI

      10.1016/j.bbrc.2017.01.165

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Directed binding of gliding bacterium, Mycoplasma mobile, shown by detachment force and bond lifetime2016

    • Author(s)
      Tanaka A, Nakane D, Mizutani M, Nishizaka T, Miyata M
    • Journal Title

      mBio

      Volume: 7 Issue: 3 Pages: 1-1

    • DOI

      10.1128/mbio.00455-16

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] F1-ATPase conformational cycle from simultaneous single-molecule FRET and rotation measurements2016

    • Author(s)
      Sugawa, M. Okazaki, K. I. Kobayashi, M. Matsui, T. Hummer, G. Masaike, T. Nishizaka, T.
    • Journal Title

      Proc Natl Acad Sci USA

      Volume: 0 Issue: 21 Pages: 0-0

    • DOI

      10.1073/pnas.1524720113

    • Related Report
      2016 Annual Research Report 2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Direct observation of rotation and steps of the archaellum in the swimming halophilic archaeon Halobacterium salinarum2016

    • Author(s)
      Kinosita, Y. Uchida, N. Nakane, D. Nishizaka, T.
    • Journal Title

      Nat Microbiol

      Volume: 1 Issue: 11 Pages: 16148-16148

    • DOI

      10.1038/nmicrobiol.2016.148

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Directed binding of gliding bacterium, Mycoplasma mobile, shown by detachment force and bond lifetime2016

    • Author(s)
      Tanaka, A. Nakane, D. Mizutani, M. Nishizaka, T. Miyata, M.
    • Journal Title

      mBio

      Volume: 0 Pages: 0-0

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Three-Dimensional Superlocalization Imaging of Gliding Mycoplasma mobile by Extraordinary Light Transmission through Arrayed Nanoholes2015

    • Author(s)
      Lee, W. Kinosita, Y. Oh, Y. Mikami, N. Yang, H. Miyata, M. Nishizaka, T. Kim, D.
    • Journal Title

      ACS Nano

      Volume: 9 Issue: 11 Pages: 10896-908

    • DOI

      10.1021/acsnano.5b03934

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Bacteriocin Protein BacL1 of Enterococcus faecalis Targets CelDivision Loci and Specifically Recognizes L-Ala2-Cross-Bridged Peptidoglycan2015

    • Author(s)
      Jun Kurushima, Daisuke Nakane, Takayuki Nishizaka, Haruyoshi Tomitaa
    • Journal Title

      Juornal of Bacteriology

      Volume: 197 Issue: 2 Pages: 286-295

    • DOI

      10.1128/jb.02203-14

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Torque Generation by Axonemal Outer-Arm Dynein2014

    • Author(s)
      Shin Yamaguchi, Kei Saito, Miki Sutoh, Takayuki Nishizaka, Yoko Y Toyoshima, Junichiro Yajima
    • Journal Title

      Biophysical Journal

      Volume: 108 Issue: 4 Pages: 872-879

    • DOI

      10.1016/j.bpj.2014.12.038

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] アーキアラとIV型線毛のモーターの動態を可視化する2017

    • Author(s)
      西坂崇之
    • Organizer
      日本物理学会 第72回年次大会
    • Place of Presentation
      大阪大学 豊中キャンパス
    • Year and Date
      2017-03-17
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Unitary steps of supermolecular motility machineries in gliding bacteria and swimming archaea2017

    • Author(s)
      Takayuki Nishizaka
    • Organizer
      Bacterial Flagella, Injectisomes and Type III Secretion Systems
    • Place of Presentation
      Okinawa Institute of Science and Technology Graduate University
    • Year and Date
      2017-03-01
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Structural and functional dynamics of nanoscale molecular motors2016

    • Author(s)
      Takayuki Nishizaka Y. Kinosita T. A. Katoh D. Nakane
    • Organizer
      31st International Congress on High Speed Imaging and Photonics
    • Place of Presentation
      Hotel Hankyu Expo Park
    • Year and Date
      2016-11-07
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Visualization of dynamics of motors in archaella and type IV pili system2016

    • Author(s)
      Takayuki Nishizaka
    • Organizer
      Current and Future Perspectives in Active Matter
    • Place of Presentation
      The University of Tokyo
    • Year and Date
      2016-10-28
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 光学顕微鏡技術の新展開:膜超分子モーターへの先進的な応用 シンポジウム「膜タンパク分子作動原理解明の新基軸」2016

    • Author(s)
      西坂崇之
    • Organizer
      第89回日本生化学会大会
    • Place of Presentation
      東北大学川内北キャンパス
    • Year and Date
      2016-09-25
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] Unitary Steps of Supermolecular Motility Machineries in Gliding Bacteria and Swimming Arcaea2016

    • Author(s)
      Kinosita, Yoshiaki Nakane, Daisuke Uchida, Nariya Miyata, Makoto Nishizaka, Takayuki
    • Organizer
      Biophyscial Society 60th Annual Meeting
    • Place of Presentation
      Los Angeles Convention Center, Los Angeles, California
    • Year and Date
      2016-02-27
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Real-time imaging of a polymorphism, rotation rate and stepwise rotation of archaeal flagella (archaella)2016

    • Author(s)
      Kinosita, Y Uchida, N Nakane, D Nishizaka, T
    • Organizer
      Sensory transduction in Microorganism (Gordon conference)
    • Place of Presentation
      Ventura Beach Marriott, Ventura, CA
    • Year and Date
      2016-01-17
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Detection of rotation of archaeal flagella under advanced optical microscopes2015

    • Author(s)
      Kinosita, Yoshiaki Nakane, Daisuke Nishizaka, Takayuki
    • Organizer
      Pacifichem 2015
    • Place of Presentation
      Honolulu, Hawaii, USA
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Mechanics of Motor Proteins and Supermolecular Motility Machinery Revealed under Advanced Optical Microscopes2015

    • Author(s)
      西坂崇之
    • Organizer
      WPI-next international workshop ~~High resolution cell biology~~
    • Place of Presentation
      E & S building, Nagoya Univ.
    • Year and Date
      2015-10-01
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] シンポジウム「べん毛・繊毛が織りなす多様な生命現象に挑む ~分子から個体まで~ Diverse functions of flagella and cilia; -From molecules to mammalian development-」 先進光学顕微鏡により明らかになる繊毛とアーキアべん毛の力学機能 Mechanical function of cilia and archaeal flagella resolved by advanced optical microscopes2015

    • Author(s)
      西坂崇之
    • Organizer
      第53回日本生物物理学会年会
    • Place of Presentation
      金沢大学 角間キャンパス 自然科学本館
    • Year and Date
      2015-09-13
    • Related Report
      2015 Annual Research Report
  • [Book] Handbook of Photonics for Biomedical Engineering2015

    • Author(s)
      Katoh, T. A. Fujimura, S. Nishizaka, T.
    • Total Pages
      15
    • Publisher
      Springer
    • Related Report
      2015 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2018-03-28  

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