Head-head coordination mechanism of kinesin during processive motility

• Project/Area Number: 24370063
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: The University of Tokyo
• Principal Investigator: TOMISHIGE Michio 東京大学, 工学(系)研究科(研究院), 准教授 (50361530)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000); Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2013: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000); Fiscal Year 2012: ¥8,710,000 (Direct Cost: ¥6,700,000、Indirect Cost: ¥2,010,000)
• Keywords: 分子モーター / 細胞内輸送 / 一分子計測（SMD） / ナノマシン / 構造変化 / 生物物理 / 一分子計測(SMD)

Outline of Final Research Achievements

Molecular motor kinesin walks along microtubules by alternately moving two motor heads, although the underlying mechanism for the head-head coordination is still unknown. Here we employed single molecule measurements with high temporal and spatial resolutions based on dark-field microscopy, and could observe intermediate states during the stepping motion of the head and the allosteric conformational change of the head. Based on these observations we showed that the direction and tension of the neck linker play pivotal roles on the head-head coordination.

Research Products

• [Journal Article] High-speed angle-resolved imaging of single gold nanorod with microsecond temporal resolution and one degree angle precision (2015)
  • Author(s): S. Enoki, R. Iino, Y. Niitani, Y. Minagawa, M. Tomishige, H. Noji
  • Journal Title: Anal. Chem. Volume: 87 Issue: 4 Pages: 2079-2086
  • DOI: 10.1021/ac502408c
  • Peer Reviewed / Open Access
• [Journal Article] CK2 activates kinesin via induction of a conformational change (2014)
  • Author(s): M. K. Mattson-Hoss, Y. Niitani, E. A. Gordon, Y. Jun, L. Bardwell, M. Tomishige, S. Gross
  • Journal Title: Proc. Natl. Acad. Sci. Volume: 111 Issue: 19 Pages: 7000-7005
  • DOI: 10.1073/pnas.1321419111
  • Peer Reviewed
• [Journal Article] Single molecule FRET observation of kinesin-1's head-tail interaction on microtubule (2013)
  • Author(s): T. Aoki, M. Tomishige, T. Ariga
  • Journal Title: BIOPHYSICS Volume: 9 Issue: 0 Pages: 149-159
  • DOI: 10.2142/biophysics.9.149
  • NAID: 130003383784
  • ISSN: 1349-2942
  • Peer Reviewed
• [Journal Article] Photoclickable Dendritic Molecular Glue : Noncovalent-to-Covalent Photochemical Transformation of Protein Hybrids (2013)
  • Author(s): Noriyuki Uchida, Kou Okuro, Yamato Niitani, Xiao Ling, Takayuki Arifja, Michio Tomishige, Takuzo Aida
  • Journal Title: Journal of the American Chemical Society Volume: 135 Issue: 12 Pages: 4684-4687
  • DOI: 10.1021/ja401059w
  • Peer Reviewed
• [Presentation] High speed single-molecule observations of the coordinated motions of kinesin motor protein (2015)
  • Author(s): Michio Tomishige
  • Organizer: Department seminar of LEBS, CNRS
  • Place of Presentation: Centre national de la recherche scientifique (France)
  • Year and Date: 2015-03-03
  • Invited
• [Presentation] 分子モーターキネシンの二足歩行運動の一分子計測 (2014)
  • Author(s): 富重道雄
  • Organizer: シンポジウム「細胞のメゾスケール構造機能」
  • Place of Presentation: 京都大学iCeMS（京都）
  • Year and Date: 2014-12-13
  • Invited
• [Presentation] Observation of the rotational motion of the motor domain during processive motility of kinesin-1 using gold nanorod (2014)
  • Author(s): 新谷大和、榎佐和子、野地博行、飯野亮太、富重道雄
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-26
• [Presentation] Single-molecule fluorescent observations of the biased binding/unbinding of the tethered kinesin head (2014)
  • Author(s): 松崎興平、富重道雄
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-25
• [Presentation] High temporal resolution observation of the kinesin head motion during processive motility (2014)
  • Author(s): Michio Tomishige
  • Organizer: Gordon Conference: Muscle & Molecular Motors
  • Place of Presentation: Mount Snow Resort, West Dover, VT (USA)
  • Year and Date: 2014-07-07
  • Invited
• [Presentation] Direct observation of binding and unbinding motions of kinesin motor domain during processive motility (2014)
  • Author(s): Michio Tomishige
  • Organizer: Tokyo ATPase Workshop
  • Place of Presentation: Takeda Hall, The University of Tokyo (Tokyo)
  • Year and Date: 2014-06-03
  • Invited
• [Presentation] Observation of the rotational motion of the motor domain during processive motility of kinesin-1 using gold nanorod (2014)
  • Author(s): Yamato Niitani, Sawako Enoki, Hiroyuki Noji, Ryota Iino, and Michio Tomishige
  • Organizer: Tokyo ATPase Workshop
  • Place of Presentation: Takeda Hall, The University of Tokyo (Tokyo)
  • Year and Date: 2014-06-02
• [Presentation] Structural basis for the coordinated processive movement of kinesin motor protein (2013)
  • Author(s): Michio Tomishige
  • Organizer: 2nd Tokyo U - Korea U Joint Workshop on Bio-Soft Matter
  • Place of Presentation: Korea University, Seol, Korea(招待講演)
  • Year and Date: 2013-03-01
• [Presentation] Direct observation of the forward stepping motion of kinesin-1 using dark-field microscopy with 50-us temporal resolution (2013)
  • Author(s): Hiroshi Isojima
  • Organizer: 57th Annual Meeting of Biophysical Society
  • Place of Presentation: Philadelphia, PA, U.S.A.
  • Year and Date: 2013-02-04
• [Presentation] Strain-dependent regulation of the kinesin-1 s catalytic activity as studied by disulfide-crosslinking of the neck linker (2013)
  • Author(s): Yamato Niitani
  • Organizer: 57th Annual Meeting of Biophysical Society
  • Place of Presentation: Philadelphia, PA, U.S.A.
  • Year and Date: 2013-02-04
• [Presentation] Direct observation of the forward stepping motion of kinesin-1 using dark-field microscopy with 50-us temporal resolution (2012)
  • Author(s): 磯島広
  • Organizer: 日本生物物理学会第50回年会
  • Place of Presentation: 名古屋大学東山キャンパス
  • Year and Date: 2012-09-22
• [Presentation] Front-head gating mechanism of kinesin-1 as studied by single molecule FRET observation of ATP binding (2012)
  • Author(s): 新谷大和
  • Organizer: 日本生物物理学会第50回年会
  • Place of Presentation: 名古屋大学東山キャンパス
  • Year and Date: 2012-09-22
• [Presentation] Single molecule FRET observation of the nucleotide-dependent conformational changes of the kinesin motor domain (2012)
  • Author(s): 小見山亮祐
  • Organizer: 日本生物物理学会第50回年会
  • Place of Presentation: 名古屋大学東山キャンパス
  • Year and Date: 2012-09-22
• [Presentation] キネシンの協調的な二足歩行運動を可能にするアロステリックな仕組み (2012)
  • Author(s): 富重道雄
  • Organizer: 第12回日本蛋白質科学会年会ワークショップ「過渡的複合体が関わる生命現象の統合的理解」
  • Place of Presentation: 名古屋国際会議場(招待講演)
  • Year and Date: 2012-06-20
• [Presentation] How kinesin coordinates two motor domains to walk along microtubule
  • Author(s): 富重道雄
  • Organizer: The 2nd KIAS Conference on Subcellular Dynamics
  • Place of Presentation: Korea Institute for Advanced Study, Seoul, Korea
  • Invited
• [Presentation] 暗視野顕微鏡によるキネシンの高時間分解能運動計測
  • Author(s): 富重道雄
  • Organizer: 第３回分子モーター討論会
  • Place of Presentation: 東大農学部中島董一郎記念ホール、東京
  • Invited
• [Presentation] Strain-dependent regulation of the kinesin-1's catalytic activity as studied by disulfide-crosslinking of the neck linker
  • Author(s): 新谷大和
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都国際会館、京都
• [Presentation] High temporal resolution observation of the stepping motion of kinesin-1 under load
  • Author(s): 穐鹿一穂
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都国際会館、京都
• [Presentation] Strain-dependent regulation of the kinesin-1's catalytic activity as studied by disulfide-crosslinking of the neck linker
  • Author(s): Yamato Niitani
  • Organizer: 58th Annual Meeting of Biophysical Society
  • Place of Presentation: San Francisco, CA, USA