Elucidating mechanisms of biological molecular machines by single-molecule physiology

• Project/Area Number: 16002013
• Research Category: Grant-in-Aid for Specially Promoted Research
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Waseda University (2005-2008); National Institutes of Natural Sciences Okazaki Research Facilities (2004)
• Principal Investigator: KINOSITA Kazuhiko Waseda University, 理工学術院, 教授 (30124366)
• Project Period (FY): 2004 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥583,700,000 (Direct Cost: ¥449,000,000、Indirect Cost: ¥134,700,000); Fiscal Year 2008: ¥110,370,000 (Direct Cost: ¥84,900,000、Indirect Cost: ¥25,470,000); Fiscal Year 2007: ¥110,890,000 (Direct Cost: ¥85,300,000、Indirect Cost: ¥25,590,000); Fiscal Year 2006: ¥112,450,000 (Direct Cost: ¥86,500,000、Indirect Cost: ¥25,950,000); Fiscal Year 2005: ¥133,510,000 (Direct Cost: ¥102,700,000、Indirect Cost: ¥30,810,000); Fiscal Year 2004: ¥116,480,000 (Direct Cost: ¥89,600,000、Indirect Cost: ¥26,880,000)
• Keywords: 一分子観察 / 一分子操作 / 回転分子モーター / F1-ATPase / リニアー分子モーター / ミオシンV / 磁気ピンセット / 光ピンセット / F_1-ATPase / トポイソメラーゼ / 歩行動作 / 回転駆動機構 / トルク / 蛍光性ATP / ATP合成酵素 / ミオシン / アクチン

Research Abstract

光学顕微鏡下の一分子生理学、とくに「一目で分かる」研究を標榜し、巨大目印による分子機械の動き・構造変化の直接可視化を通じて、動作機構の解明を試みた。回転分子モーターF1-ATPaseにおける化学反応→力学的仕事の共役スキームをほぼ完成させ、分子機械一般に通じ得る共役原理を提出した。一方、回転軸無しでも回転するという意外な発見をし、構造に基づく機能の説明は振出しに戻った。二本足のモーター、ミオシンの脚の動きの直接観察に成功し、ブラウン運動をうまく使って歩くことを証明した。

Research Products

• [Journal Article] A giant liposome for single-molecule observation of conforma-tional changes in membrane proteins (2009)
  • Author(s): Y. Onoue, T. Suzuki, M. Davidson, M. Karlsson, O. Orwar, M. Yoshida, and K. Kinosita Jr
  • Journal Title: Biochim. Biophys. Acta (Biomembranes) 1788 Pages: 1332-1340
  • Peer Reviewed
• [Journal Article] A giant liposome for single-molecule observation of conformational changes in membrane proteins (2009)
  • Author(s): Yasuhiro Onoue, et.al.
  • Journal Title: Biochimica et Biophysica Acta (BBA)-Biomembranes 1788 Pages: 1332-1340
  • Peer Reviewed
• [Journal Article] Neither helix in the coiled coil region of the axle of F1-ATPase plays a significant role in torque production (2008)
  • Author(s): M. D. Hossain, S. Furuike, Y. Maki, K. Adachi, T. Suzuki, A. Kohori, H. Itoh, M. Yoshida & K. Kinosita Jr
  • Journal Title: Biophys. J. 95 Pages: 4837-4844
  • Peer Reviewed
• [Journal Article] Temperature dependence of the rotation and hydrolysis activities of F1-ATPase (2008)
  • Author(s): S. Furuike, K. Adachi, N. Sakaki, R. Shimo-Kon, H. Itoh, E. Muneyuki, M. Yoshida & K. Kinosita Jr
  • Journal Title: Biophys. J. 95 Pages: 761-770
  • Peer Reviewed
• [Journal Article] Axle-less F1-ATPase rotates in the correct direction (2008)
  • Author(s): S. Furuike, M. D. Hossain, Y. Maki, K. Adachi, T. Suzuki, A. Kohori, H. Itoh, M. Yoshida & K. Kinosita Jr
  • Journal Title: Science 319 Pages: 955-958
  • Peer Reviewed
• [Journal Article] Neither helix in the coiled coil region of the axle of F1-ATPase plays a significant role in torque production (2008)
  • Author(s): Mohammad Delawar Hossain, et.al.
  • Journal Title: Biophysical Journal 95 Pages: 4837-4844
  • Peer Reviewed
• [Journal Article] Temperature dependence of the rotation and hydrolysis activities of F1-ATPase (2008)
  • Author(s): Shou Furuike, et.al.
  • Journal Title: Biophysical Journal 95 Pages: 761-770
  • Peer Reviewed
• [Journal Article] Axle-less F1-ATPase rotates in the correct direction (2008)
  • Author(s): Shou Furuike, et.al.
  • Journal Title: Science 319 Pages: 955-958
  • Peer Reviewed
• [Journal Article] Axle-less F1-ATPase rotates in the correct direction. (2008)
  • Author(s): Shou Furuike, et al.
  • Journal Title: Science 319 Pages: 955-958
• [Journal Article] Coupling of rotation and catalysis in F1-ATPase revealed by single-molecule imaging and manipulation (2007)
  • Author(s): K. Adachi, K. Oiwa, T. Nishizaka, S. Furuike, H. Noji, H. Itoh, M. Yoshida & K. Kinosita, Jr
  • Journal Title: Cell 130 Pages: 309-321
  • Peer Reviewed
• [Journal Article] Myosin V walks by lever action and Brownian motion (2007)
  • Author(s): K. Shiroguchi & K. Kinosita, Jr
  • Journal Title: Science 316 Pages: 1208-1212
  • Peer Reviewed
• [Journal Article] On the walking mechan- ism of linear molecular motors (2007)
  • Author(s): K. Kinosita, Jr., K. Shiroguchi, M. Y. Ali, K. Adachi, & H. Itoh
  • Journal Title: Adv. Exp. Med. Biol. 592 Pages: 369-384
• [Journal Article] Coupling of rotation and catalysis in F1-ATPase revealed by single-molecule imaging and manipulation. (2007)
  • Author(s): Kengo Adachi, et al.
  • Journal Title: Cell 130 Pages: 309-321
• [Journal Article] Myosin V walks by lever action and Brownian motion. (2007)
  • Author(s): Katsuyuki Shiroguchi, and Kazuhiko Kinosita, Jr.
  • Journal Title: Science 316 Pages: 1208-1212
• [Journal Article] Visualization of RecA filaments and DNA by fluorescence microscopy (2007)
  • Author(s): Taro Nishinaka, et al.
  • Journal Title: J. Biochem 141 Pages: 147-156
• [Journal Article] The rotor tip inside a bearing of a thermophilic F1-ATPase is dispensable for torque generation (2006)
  • Author(s): M. D. Hossain, S. Furuike, Y. Maki, K. Adachi, M. Y. Ali, M. Huq, H. Itoh, M. Yoshida & K. Kinosita Jr
  • Journal Title: Biophys. J. 90 Pages: 4195-4203
  • Peer Reviewed
• [Journal Article] The rotor tip inside a bearing of a thermophilic F_1-ATPase is dispensable for torque generation (2006)
  • Author(s): Mohammad Delawar Hossain, et al.
  • Journal Title: Biophys. J 90 Pages: 4195-4203
• [Journal Article] On the walking mechanism of linear molecular motors (2006)
  • Author(s): Kazuhiko Kinosita, Jr., et al.
  • Journal Title: Adv. Exp. Med. Biol. 592 Pages: 369-384
• [Journal Article] One rotary mechanism for F1-ATPase over ATP concentrations from millimolar down to nanomolar (2005)
  • Author(s): N. Sakaki, R. Shimo-Kon, K. Adachi, H. Itoh, S. Furuike, E. Muneyuki, M. Yoshida & K. Kinosita Jr
  • Journal Title: Biophys. J. 88 Pages: 2047-2056
  • Peer Reviewed
• [Journal Article] How two-foot molecular motors may walk (2005)
  • Author(s): K. Kinosita, Jr., M. Y. Ali, K. Adachi, K. Shiroguchi, & H. Itoh
  • Journal Title: Adv. Exp. Med. Biol. 565 Pages: 205-219
• [Journal Article] How two-foot molecular motors may walk (2005)
  • Author(s): Kazuhiko Kinosita, Jr., et al.
  • Journal Title: Adv.Exp.Med.Biol. 565 Pages: 205-219
• [Journal Article] Is your vide camera linear? (2005)
  • Author(s): Kazuhiko Kinosita, Jr., et al.
  • Journal Title: Seibutsu Butsuri (Biophysics, in Japanese) 45 Pages: 216-218
• [Journal Article] One rotary mechanism for F_1-ATPase over ATP concentrations from millimolar down to nanomolar (2005)
  • Author(s): Sakaki, N., Shimo-Kon, R., Adachi, K., Itoh, H., Furuike, S., Muneyuki, E., Yoshida, M., Kinosita, K.Jr.
  • Journal Title: Biophys.J. 88 Pages: 2047-2056
• [Journal Article] ATP-driven stepwise rotation F_0F_1-ATP synthase (2005)
  • Author(s): Ueno, H., Suzuki, T., Kinosita, K.Jr., Yoshida, M.
  • Journal Title: Proc.Natl.Acad.Sci.USA 102 Pages: 1333-1338
• [Journal Article] Activation of pausing F_1 motor by external force (2005)
  • Author(s): Hirono-Hara, Y., Ishizuka, K., Kinosita, K.Jr., Yoshida, M., Noji, H.
  • Journal Title: Proc.Natl.Acad.Sci.USA 102 Pages: 4288-4293
• [Journal Article] Unconstrained steps of myosin VI appear longest among known molecular motors (2004)
  • Author(s): M. Y. Ali, K. Homma, A. H. Iwane. K. Adachi, H. Itoh, K. Kinosita Jr. T. Yanagida, & M. Ikebe
  • Journal Title: Biophys. J. 86 Pages: 3804-3810
  • Peer Reviewed
• [Journal Article] Rotation of F1-ATPase : How an ATP-driven molecular machine may work (2004)
  • Author(s): K. Kinosita, Jr., K. Adachi, & H. Itoh
  • Journal Title: Annu. Rev. Biophys. Biomol. Struct. 33 Pages: 245-268
  • Peer Reviewed
• [Journal Article] Unconstrained steps of myosin VI appear longest among known molecular motors (2004)
  • Author(s): Ali, M.Y., Homma, K., Iwane, A.H., Adachi, K., Itoh, H., Kinosita, K.Jr., Yanagida, T., Ikebe, M.
  • Journal Title: Biophys.J. 86 Pages: 3804-3810
• [Journal Article] Rotation of F_1-ATPase : How an ATP-driven molecular machine may work (2004)
  • Author(s): Kinosita, K.Jr., Adachi, K., Itoh, H.
  • Journal Title: Annu.Rev.Biophys.Biomol.Struct. 33 Pages: 245-268
• [Journal Article] Temperature Dependence of the Rotation and Hydrolysis Activities of F_1-ATPase
  • Author(s): Shou Furuike, et al.
  • Journal Title: Biophysical Journal (in press)
• [Journal Article] The rotor tip inside a bearing of a thermophilic F_1-ATPase is dispensable for torque generation
  • Author(s): Mohammad Delawar Hossain, et al.
  • Journal Title: Biophys.J. (in press)
• [Journal Article] On the walking mechanism of linear molecular motors
  • Author(s): Kazuhiko Kinosita, Jr., et al.
  • Journal Title: Adv.Exp.Med.Biol. (in press)
• [Presentation] II型DNAトポイソメラーゼの手品のような反応の一分子解析 (2009)
  • Author(s): 余語克紀, 他
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 東京大学, 東京
• [Presentation] TFoFlのATP合成 (2009)
  • Author(s): 曽我直樹, 他
  • Organizer: 第43回生体運動合同班会議
  • Place of Presentation: 東京大学, 東京
• [Presentation] ミオシンVの歩き方 : 浮いた足(もと後足)が前方に着地できる仕組み (2008)
  • Author(s): 城口克之
  • Organizer: 早稲田大学理工学術院物理学科高野光則研究室
  • Place of Presentation: 早稲田大学, 東京
  • Year and Date: 2008-06-30
• [Presentation] "How does F1-ATPase rotate? "Nobel Symposium on single molecule spectroscopy in chemistry, physics and biology (2008)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited symposium lecture
  • Place of Presentation: Stockholm, Sweden
• [Presentation] How does F1-ATPase rotate? (2008)
  • Author(s): Kazuhiko Kinosita, Jr.
  • Organizer: Nobel Symposium on Single molecule spectroscopy in chemistry, physics and biology
  • Place of Presentation: Stockholm, Sweden
• [Presentation] Watching protein motors to work. (2008)
  • Author(s): Kazuhiko Kinosita, Jr.
  • Organizer: Complex Nanosystems : Assembly, Control and Functionality
  • Place of Presentation: San Servolo, Italy
• [Presentation] Protein Machines under an Optical Microscope (2008)
  • Author(s): Kazuhiko Kinosita, Jr.
  • Organizer: The 39th NIPS International Symposium. Frontiers of Biological Imaging. Synergy of the Advanced Techniques
  • Place of Presentation: Okazaki, Japan
• [Presentation] 軸なしF_1-ATPaseも正しい方向へ回転する (2008)
  • Author(s): 古池晶, 他
  • Organizer: 第8回日本蛋白質科学会年会
  • Place of Presentation: タワーホール船堀, 東京
• [Presentation] ATP駆動の回転において塾ATPaseは特徴的なステップ状回転を示す (2008)
  • Author(s): 古池晶, 他
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場, 福岡
• [Presentation] F_1-ATPase回転モーターの角度依存的ヌクレオチドアフィニティ"Angle Dependence of the Afanity for Nucleotides in Rotary Motor F_1-ATPase" (2008)
  • Author(s): 足立健吾, 他
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場,福岡
• [Presentation] リバースジャイレースの一分子解析-正の超らせんをつくるトポイソメラーゼ (2008)
  • Author(s): 小川泰策, 他
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場,福岡
• [Presentation] Effects of sodium dodecyl sulfate on F_1-ATPase (2008)
  • Author(s): Mohammad Delawar Hossain, et.al.
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場,福岡
• [Presentation] II型トポイソメラーゼが手品のようにDNAの絡まりを解くところを一分子操作・観察する (2008)
  • Author(s): 余語克紀, 他
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡国際会議場, 福岡
• [Presentation] ミオシンVの足首の動きを見る(アクチンなし) (2008)
  • Author(s): 城口克之, 他
  • Organizer: 日本生物物理学会第46回年会
  • Place of Presentation: 福岡国際会議場, 福岡
• [Presentation] Single-molecule observation of unlinking magic by type-II topoisomerases with real-time control of DNA substrates (2008)
  • Author(s): 余語克紀, 他
  • Organizer: BMB2008(第31回日本分子生物学会年会、第81回日本生化学会年会合同)
  • Place of Presentation: 神戸ポートアイランド, 神戸
• [Presentation] F_1-ATPase with fused γ and β subunits. (2008)
  • Author(s): Ayako Kohori, Katsuyuki Shiroguchi, Shou Furuike, Masasuke Yoshida, Kazuhiko Kinosita, Jr
  • Organizer: 16th IUPAB International Biophysics Congress.
  • Place of Presentation: Long Beach, California, U.S.A.
• [Presentation] Direct observation of DNA untangling magic by a type-II top oisomerase. (2008)
  • Author(s): Katsunori Yogo, Taisuke Ogawa, Kazuhiko Kinosita, Jr
  • Organizer: 16th IUPAB International of Biophysics Congress.
  • Place of Presentation: Long Beach, California, U.S.A.
• [Presentation] DNAトポイソメラーゼIIの1分子解析 (2008)
  • Author(s): 小川泰作、余語克紀、小畠紗季、木下一彦
  • Organizer: 2008年 生体運動研究合同班会議
  • Place of Presentation: 仙台市戦災復興記念館記念ホール
• [Presentation] DNAトポイソメラーゼIIが絡まりを解くところを観る。 (2008)
  • Author(s): 余語克紀、小川泰作、小畠紗季、木下一彦
  • Organizer: 2008年生体運動研究合同班会議
  • Place of Presentation: 仙台市戦災復興記念会記念ホール
• [Presentation] "Single-molecule physiology of protein machines" European Biophysics Congress (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited plenary lecture
  • Place of Presentation: London, England
• [Presentation] "Single-molecule physiology under an optical microscope" XXIII Biennial of the Society of Microscopy of Spain (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited plenary lecture
  • Place of Presentation: Bilbao, Spain
• [Presentation] "Single-molecule physiology of protein motors" Nano Tech Insight (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited plenary lecture
  • Place of Presentation: Luxor, Egypt
• [Presentation] Single-molecule physiology under an optical microscope. (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: XXIII Biennial of the Society of Microscopy of Spain
  • Place of Presentation: Spain.
• [Presentation] Single-molecule physiology of protein machines. (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: European Biophysics Congress London 2007
  • Place of Presentation: London, England
• [Presentation] How molecular machines may sense the free energy? (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: The Third Workshop of the UK-Japan Bionanotechnology Collaboration.
  • Place of Presentation: Oxford, England
• [Presentation] Single-molecule physiology of protein machines. (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: 30th Anniversary Symposium of Institute of Biological Chemistry (IBC), Academia Sinica Proteins-from Chemistry to Biology
  • Place of Presentation: Taipei, Taiwan.
• [Presentation] How dose F_1ATPase rotate? (2007)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: Engineering Life
  • Place of Presentation: Germany.
• [Presentation] トポイソメラーゼIIがDNAの絡まりを解くところを観る (2007)
  • Author(s): 余語克紀、小川泰作、木下一彦
  • Organizer: Biochemistry and Molecular Biology
  • Place of Presentation: パシフィコ横浜
• [Presentation] F_1-ATPaseのATP加水分解中のヌクレオチド結合数、非触媒部位にヌクレオチドが結合できない異変体を使って (2007)
  • Author(s): 下-昆理恵子、宗行英朗、足立健吾、古池晶、酒井坦、吉田賢右、木下一彦
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 吊られたアクチンフィラメント上の束縛されないミオシンVの運動の直接観察 (2007)
  • Author(s): 田島勇治、城口克之、木下一彦
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] マイクロビーズがF1-ATPaseの回転方向と逆に回転する現象について(P) (2007)
  • Author(s): 木下昌威、木下一彦
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] γ-β fusion mutantを用いたF1-ATPaseの回転におけるγサブユニットN末端の役割の解明(P) (2007)
  • Author(s): 小堀綾子、城口克之、古池晶、吉田賢右、木下一彦
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Studies of subunit-truncated F_1-ATPase. (2007)
  • Author(s): Delawar Mohammad Hossain, 古池晶、牧泰史、足立健吾、木下一彦
  • Organizer: 第45回日本生物物理学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] "Probing Nature's Nanoscale Machines with Micro- scale Probes" Biophysical Society 50th annual meeting (2006)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited lecture as the 2006 National Lecturer
  • Place of Presentation: Salt Lake City, Utah, USA
• [Presentation] "F1-ATPase : A molecular trans- ducer of chemical and mechanical energies" Nobel Symposium 132 : Energy in Cosmos, Molecules and Life (2005)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited symposium lecture
  • Place of Presentation: Stockholm, Sweden
• [Presentation] "F1-ATPase : a rotary motor / generator" Nobel Symposium 131 : Controlled Nanoscale Motion in Biological and Artificial Systems (2005)
  • Author(s): Kazuhiko Kinosita, Jr
  • Organizer: invited symposium lecture
  • Place of Presentation: Backaskog, Sweden
• [Remarks] 研究室ホームページ(業績他)
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• [Remarks]
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