Demonstration of molecular machine by control of microtubule rigidity and nano-patterning

• Project/Area Number: 17H03206
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Intelligent mechanics/Mechanical systems
• Research Institution: Kyoto University
• Principal Investigator: Yokokawa Ryuji 京都大学, 工学研究科, 教授 (10411216)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000); Fiscal Year 2019: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2018: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000); Fiscal Year 2017: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
• Keywords: 分子ロボティクス / マイクロ・ナノデバイス / 生物物理 / 分子モーター / ナノバイオ

Outline of Final Research Achievements

The aim was to develop a design methodology of kinesin motor-driven molecular machine by controlling flexural rigidity of microtubules and spacing of motors. Flexural rigidity was measured by analyzing a fluctuating microtubule immobilized on a glass substrate. We found the growth rate of microtubules has significant influence on the rigidity compared with microtubule associated proteins and the difference of nucleotides. In addition, we found the different motility of two kinesins immobilized on gold nano pillars.

Academic Significance and Societal Importance of the Research Achievements

本研究成果は、微小管の曲げ剛性と生体分子モーターの配置間隔に着目して、ナノマイクロ加工技術とモーターを利用した分子機械の設計論を体系化することにつながるものである。この過程において、微小管の曲げ剛性が伸張速度により支配的に決まることや、モーターの種類によって運動特性が異なることを見出した。これらは、分子機械の開発と言うナノマイクロ工学的視点から研究を進めることにより得られた新たな知見であるが、同時に生物物理学における中心的な課題でもあり当該分野への貢献も大きい。

Research Products

• [Int'l Joint Research] University of Massachusetts/University of Wisconsin(米国)
• [Int'l Joint Research] University of Massachusetts/University of Wisconsin(米国)
• [Int'l Joint Research] University of Massachusetts/University of Wisconsin(米国)
• [Journal Article] Different motilities of microtubules driven by kinesin-1 and kinesin-14 motors patterned on nanopillars. (2020)
  • Author(s): Taikopaul Kaneko,Ken'ya Furuta,Kazuhiro Oiwa,Hirofumi Shintaku,Hidetoshi Kotera,Ryuji Yokokawa
  • Journal Title: Science Advances Volume: 6 Issue: 4 Pages: 7413-7413
  • DOI: 10.1126/sciadv.aax7413
  • NAID: 120006779802
  • Peer Reviewed / Open Access
• [Journal Article] Spatial Patterning of Kinesin-1 and Dynein Motor Proteins in an In Vitro Assay using Aqueous Two-Phase Systems (ATPS). (2019)
  • Author(s): Tamanna Ishrat Farhana,Tomohiro Nakagawa,Shumpei Ohara,Hirofumi Shintaku,Hidetoshi Kotera,Ryuji Yokokawa
  • Journal Title: Langmuir Volume: 35 Issue: 40 Pages: 13003-13010
  • DOI: 10.1021/acs.langmuir.9b01411
  • Peer Reviewed
• [Journal Article] Microtubule Polymerization in Alignment by an On-Chip Teperature Gradient Platform (2019)
  • Author(s): J.-H.Liu,K.-C. Hsia,R.Yokokawa,Y.-W.Lu
  • Journal Title: Sens. Actuator, B. Volume: 298 Pages: 126813-126813
  • DOI: 10.1016/j.snb.2019.126813
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transport of microtubules according to the number and spacing of kinesin motors on gold nano-pillars (2019)
  • Author(s): Kaneko Taikopaul、Ando Suguru、Furuta Ken’ya、Oiwa Kazuhiro、Shintaku Hirofumi、Kotera Hidetoshi、Yokokawa Ryuji
  • Journal Title: Nanoscale Volume: 000 Issue: 20 Pages: 1-9
  • DOI: 10.1039/c9nr01324e
  • Peer Reviewed
• [Journal Article] Characterization of Microtubules Gliding on Surfaces Roughness Structure (2018)
  • Author(s): 中原 佐,宮﨑 勇,小寺 秀俊,横川 隆司,南 和幸
  • Journal Title: IEEJ Transactions on Sensors and Micromachines Volume: 138 Issue: 11 Pages: 503-508
  • DOI: 10.1541/ieejsmas.138.503
  • NAID: 40021726562
  • ISSN: 1341-8939, 1347-5525
  • Year and Date: 2018-11-01
  • Peer Reviewed
• [Journal Article] Simultaneous observation of kinesin-driven microtubule motility and binding of adenosine triphosphate using linear zero-mode waveguides (2018)
  • Author(s): Fujimoto K, Morita Y, Iino R, Tomishige M, Shintaku H, Kotera H, Yokokawa R
  • Journal Title: ACS Nano Volume: 12 Issue: 12 Pages: 11975-11985
  • DOI: 10.1021/acsnano.8b03803
  • Peer Reviewed
• [Journal Article] 分子を狙いどおりに目的地へ送り込む!―分子機械と微小流体デバイスによるナノ輸送システム― (2018)
  • Author(s): 磯崎直人,横川隆司
  • Journal Title: 化学 Volume: 73
  • NAID: 40021444453
• [Journal Article] Control of molecular shuttles by designing electrical and mechanical properties of microtubules (2017)
  • Author(s): Isozaki Naoto、Shintaku Hirofumi、Kotera Hidetoshi、Hawkins Taviare L.、Ross Jennifer L.、Yokokawa Ryuji
  • Journal Title: Science Robotics Volume: 2 Issue: 10
  • DOI: 10.1126/scirobotics.aan4882
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Design and Fabrication of Linear-shaped Zero Mode Waveguides for Single Molecule Observation of Kinesin and Fluorescent ATP (2017)
  • Author(s): Fujimoto Kazuya、Morita Yuki、Iino Ryota、Tomishige Michio、Shintaku Hirofumi、Kotera Hidetoshi、Yokokawa Ryuji
  • Journal Title: IEEJ Transactions on Sensors and Micromachines Volume: 137 Issue: 6 Pages: 159-164
  • DOI: 10.1541/ieejsmas.137.159
  • NAID: 130005687854
  • ISSN: 1341-8939, 1347-5525
  • Peer Reviewed
• [Journal Article] Single Molecule Manipulation: Pick-and-Place Assembly of Single Microtubules (Small 32/2017) (2017)
  • Author(s): Tarhan Mehmet Cagatay、Yokokawa Ryuji、Jalabert Laurent、Collard Dominique、Fujita Hiroyuki
  • Journal Title: Small Volume: 13 Issue: 32 Pages: 1701136-1701136
  • DOI: 10.1002/smll.201770172
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Microtubule density and landing rates as parameters to analyze tau protein in the MT-kinesin “gliding” assay (2017)
  • Author(s): S. Subramaniyan Parimalam, M. C. Tarhan, S. L. Karsten, H. Fujita, H. Shintaku, H. Kotera, R. Yokokawa
  • Journal Title: Sens. Actuators, B Volume: 238 Pages: 954-964
  • DOI: 10.1016/j.snb.2016.07.082
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] Molecular Nano-Patterning Reveals Different Coordination of Kinesin-1 and Kinesin-14 Motors (2020)
  • Author(s): T. Kaneko,K. y. Furuta,K. Oiwa,H. Shintaku,H. Kotera,R. Yokokawa
  • Organizer: 64th Annual Meeting of Biophysical Society
  • Int'l Joint Research
• [Presentation] Motility of Microtubule Driven by Kinesin in a Viscoelastic Media on Au Nano-Pillar Substrate (2020)
  • Author(s): M. Furukawa,T. Kaneko,R. Yokokawa
  • Organizer: The 33rd International Conference on Micro Electro Mechanical Systems (MEMS 2020)
  • Int'l Joint Research
• [Presentation] The Cooperative Motility of Microtubules on Nano-Patterned Kinesin-1 TURF (2019)
  • Author(s): T. I. Farhana, T. Kaneko, R. Yokokawa
  • Organizer: The 20th International Conference on Solid-State Sensors, Actuators and Microsystems (Transducers 2019)
  • Int'l Joint Research
• [Presentation] Patterning of Different Motor Proteins Using Aqueous Two-Phase (2019)
  • Author(s): T. Nakagawa, S. Oohara, T. I. Farhana, R. Yokokawa
  • Organizer: The 20th International Conference on Solid-State Sensors, Actuators and Microsystems (Transducers 2019)
  • Int'l Joint Research
• [Presentation] Investigation of Collisions of Microtubules Driven by Nano-Patterned Kinesins (2019)
  • Author(s): T. I. Farhana, T. Kaneko, R. Yokokawa
  • Organizer: 63rd Annual Meeting of Biophysical Society
  • Int'l Joint Research
• [Presentation] Flexural Rigidity of Microtubules Measured with Nanometer-Level Localization Precision (2019)
  • Author(s): H. Zhou, N. Isozaki, T. L. Hawkins, J. L. Ross, R. Yokokawa
  • Organizer: 63rd Aunnal Meeting of Biophysical Society
  • Int'l Joint Research
• [Presentation] 粘弾性溶液中におけるキネシンによる微小管双方向運動の解析 (2019)
  • Author(s): 古川 眞之, 金子 泰洸ポール, Tamanna Ishrat Farhana, 横川 隆司
  • Organizer: 第36回「センサ・マイクロマシンと応用システム」シンポジウム
• [Presentation] 分子パターニング技術を用いた微小管すれ違い現象の解析 (2019)
  • Author(s): 東條 裕也, 金子 泰洸ポール, 古田 健也, 大岩 和弘, 横川 隆司
  • Organizer: 第40回化学とマイクロナノシステム研究会
• [Presentation] The Effect of Microtubules’ Flexural Rigidity on Their Collective Motion (2019)
  • Author(s): H. Zhou, Y. Tojo, T. I. Farhana, R. Yokokawa
  • Organizer: 第40回化学とマイクロナノシステム研究会
• [Presentation] Study of Collisions of Microtubules on Nano-Patterned Kinesin Surface (2019)
  • Author(s): T. I. Farhana, T. Kaneko, R. Yokokawa
  • Organizer: 第40回化学とマイクロナノシステム研究会
• [Presentation] Emergence of Collective Motion of Microtubules Assisted by Spatial Arrangements of Kinesin-1 (2019)
  • Author(s): Tamanna Farhana Ishrat, Taikopaul Kaneko, Ryuji Yokokawa
  • Organizer: 第39回化学とマイクロナノシステム研究会
• [Presentation] The Influence of 2N4R Tau Isoform on the Flexural Rigidity of Microtubule Revealed through Nanometer-level Localization Precision (2019)
  • Author(s): Hang Zhou, Kosuke Naka, Taviare L Hawkins, Jennifer L Ross, Ryuji Yokokawa
  • Organizer: 第39回化学とマイクロナノシステム研究会
• [Presentation] 1分子ナノパターニングによるkinesin-1およびncdの協働性の評価 (2019)
  • Author(s): 金子 泰洸ポール, 大庭 将太郎, 古田 健也, 大岩 和弘, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 第39回化学とマイクロナノシステム研究会
• [Presentation] 粘弾性溶液中における微小管の双方向運動の観察 (2019)
  • Author(s): 古川 眞之, 金子 泰洸ポール, Farhana Tamanna Ishrat, 横川 隆司
  • Organizer: 第39回化学とマイクロナノシステム研究会
• [Presentation] Micro/Nano Fabrications for Biophysical Studies of Motor Proteins and Organ-on-a-Chip Applications (2018)
  • Author(s): R. Yokokawa
  • Organizer: JSPS-RS Joint Symposium
  • Int'l Joint Research / Invited
• [Presentation] Micro/Nano Fabrications for Biophysics and Organ-on-a-Chip Applications (2018)
  • Author(s): R. Yokokawa
  • Organizer: IEEE Electron Devices Technology and Manufacturing (EDTM) Conference
  • Int'l Joint Research / Invited
• [Presentation] Investigating Coordination of Kinesin Motor Proteins using Their Selective Immobilization on Gold Nano-Pillars (2018)
  • Author(s): T. Kaneko, S. Ohba, K. Furuta, K. Oiwa, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: The 22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences (?TAS 2018)
  • Int'l Joint Research
• [Presentation] Development of a Nano-Patterning of Kinesin to Control the Number and Arrangement of Motors by Combining Au Nano-Pillars and SAM (2018)
  • Author(s): T. Kaneko, S. Ohba, K. Furuta, K. Oiwa, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: Molecules, Materials, Devices and Systems in Medicine
  • Int'l Joint Research
• [Presentation] Different Coordination of Kinesin-1 and Ncd Revealed by Their Selective Immobilization on Gold Nano-Pillars (2018)
  • Author(s): T. Kaneko, S. Ohba, K. Furuta, K. Oiwa, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: Biophysical Society 62nd Annual Meeting
  • Int'l Joint Research
• [Presentation] Fluorescent Observation of ATP Binding in Kinesin Driven Microtubule Gliding Using Nano-Slits (2018)
  • Author(s): K. Fujimoto, R. Iino, M. Tomishige, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: The 31st IEEE International Conference on Micro Electro Mechanical Systems (MEMS 2018)
  • Int'l Joint Research
• [Presentation] ATPSによる異種モータタンパク質の固定と運動性への影響の評価 (2018)
  • Author(s): 中川 倫宏, 大原 駿平, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 第38回化学とマイクロナノシステム研究会
• [Presentation] キネシンの1分子ナノパターニングによる微小管の運動方向の制御 (2018)
  • Author(s): 金子 泰洸ポール, 大庭 将太郎, 古田 健也, 大岩 和弘, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 第38回化学とマイクロナノシステム研究会
• [Presentation] Relationship between Microtubule's Flexural Rigidity and Growth Rate by Controlling Tubulin Concentration (2018)
  • Author(s): Zhou Hang, Isozaki Naoto, Hawkins Taviare L., Ross Jennifer L., Yokokawa Ryuji
  • Organizer: 第38回化学とマイクロナノシステム研究会
• [Presentation] Regulating the Collective Motility of Microtubules by Controlling the Spatial Arrangement of Kinesins (2018)
  • Author(s): Tamanna Ishrat Farhana; Taikopaul Kaneko Ryuji Yokokawa
  • Organizer: The 35th Sensor Symposium on Sensors, Micromachines and Applied Systems
• [Presentation] Selective Nano-Patterning of Kinesin Motor-Proteins and Its Effect on Collective Motion of Microtubules (2018)
  • Author(s): Farhana Tamanna Ishrat, Kaneko Taikopaul, Yokokawa Ryuji
  • Organizer: 56th Annual Meeting of Biophysical Society Japan
• [Presentation] 粘弾性溶液中におけるキネシンによる微小管の運動についての研究 (2018)
  • Author(s): 古川 眞之, 金子 泰洸ポール, Farhana Tamanna Ishrat, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 第56回日本生物物理学会年会
• [Presentation] Investigating Coordination of Kinesin-1 and Ncd Using Their Selective Immobilization on Gold Nano-Pillars (2018)
  • Author(s): 金子 泰洸ポール, 大庭 将太郎, 古田 健也, 大岩 和弘, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 第56回日本生物物理学会年会
• [Presentation] 分子パターニングを用いたモータタンパク質集団の同期現象の解明 (2018)
  • Author(s): 金子 泰洸ポール, 大庭 将太郎, 古田 健也, 大岩 和弘, 新宅 博文, 小寺 秀俊, 横川 隆司
  • Organizer: 西日本ナノシート研究会サマーキャンプ2018
• [Presentation] 微小管の曲げ剛性評価ため全反射顕鏡による伸長速度測定 (2018)
  • Author(s): 浮田 一輝, Zhou Hang, 新宅 博文, 小寺 秀俊, Hawkins Taviare L., Ross Jennifer L., 横川 隆司
  • Organizer: 電気学会_バイオマイクロシステム研究会
• [Presentation] マイクロチャネル内におけるATPS を用いたモータタンパク質のパターニング (2018)
  • Author(s): 中川倫宏 新宅博文, 小寺秀俊, 横川隆司
  • Organizer: 電気学会全国大会
• [Presentation] Different coordination of kinesin-1 and Ncd revealed by their selective immobilization on gold nano-pillars (2017)
  • Author(s): T. Kaneko, S. Ohba, K. Furuta, K. Oiwa, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: Biophysical Society 62nd Annual Meeting
  • Int'l Joint Research
• [Presentation] Fluorescent observation of ATP binding in kinesin driven microtubule gliding using nano-slits (2017)
  • Author(s): K. Fujimoto, R. Iino, M. Tomishige, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: The 31st IEEE International Conference on Micro Electro Mechanical Systems (MEMS 2018)
  • Int'l Joint Research
• [Presentation] Microtubule Polymerization Temperature Gradient in Microchip (2017)
  • Author(s): J. H. Liu, R. Yokokawa, Y. W. Lu
  • Organizer: The 21st International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS 2017)
  • Int'l Joint Research
• [Presentation] High-Precision Microtubule Sorting by the Measurement of Persistence Length with Sub-Pixel Resolution (2017)
  • Author(s): N. Isozaki, K. Ukita, H. Shintaku, H. Kotera, T. L. Hawkins, J. L. Ross, R. Yokokawa
  • Organizer: 2017 BMES Annual Meeting
  • Int'l Joint Research
• [Presentation] Integration of Au Nano-Pillars and Sam Enables Protein Patterning with Designed Spacing at Single Molecule Level (2017)
  • Author(s): T. Kaneko, K. Sasakura, K. Furuta, K. Oiwa, H. Shintaku, H. Kotera, R. Yokokawa
  • Organizer: The17th IEEE International Conference on Nanotechnology (IEEE NANO 2017)
  • Int'l Joint Research
• [Presentation] Micro/nano fabrications for biophysical studies of endothelial cells and motor proteins (2017)
  • Author(s): R. Yokokawa
  • Organizer: International Conference on Molecular Electronics and Bioelectronics (M&BE9)
  • Int'l Joint Research / Invited
• [Remarks] 研究室HP
  • URL: http://www.ksys.me.kyoto-u.ac.jp/