Construction of an atomic model of the skeletal muscle thin filament by FRET measurements

• Project/Area Number: 21570162
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: University of Fukui
• Principal Investigator: MIKI Masao 福井大学, 大学院・工学研究科, 教授 (30242580)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2009: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: タンパク質 / 核酸の構造 / 動態 / 機能 / アクチン / トロポミオシン / トロポニン / 筋収縮制御 / FRET / 細いフィラメント / カルシウム制御 / アトミックモデル / 生物物理 / 分子スイッチ / 蛍光測定 / 制御メカニズム

Research Abstract

Striated muscle thin filament is composed of actin, tropomyosin(Tm), and troponin(Tn). The fluorescence resonance energy(FRET) donor probe was labeled at the specific sites on tropomyosin or troponin, and the energy acceptor probes were labeled at the specific sites on actin. From these donor and acceptor pairs, FRET efficiencies were determined. Using the atomic coordinates of F-actin, Tm, and Tn, we searched all possible arrangements for Tn and Tm on F-actin to calculate FRET efficiency for each donor-acceptor pair in each arrangement. By minimizing the squared sum of deviations for the calculated FRET efficiencies from the observed efficiencies, we determined the location of Tm and Tn on F-actin.

Research Products

• [Journal Article] A Three-Dimensional FRET Analysis to Construct an Atomic Model of the Actin. Tropomyosin. Troponin Core Domain Complex on a Muscle Thin Filament (2012)
  • Author(s): Miki, M., Makimura, S., Sugahara, Y., Yamada, R., Bunya, M., Saitoh, T., & Tobita, H.
  • Journal Title: J. Mol. Biol Volume: 420 Pages: 40-55
  • NAID: 120004186302
  • Peer Reviewed
• [Journal Article] A Three Dimensional FRET Analysis to construct an Atomic Model of the Actin-Tropomyosin-Troponin Core Domain Complex on a Muscle Thin Filament (2012)
  • Author(s): Miki, M., Makimura, S., Sugahara, Y., Yamada, R., Bunya, M., Saitoh, T., Tobita, H.
  • Journal Title: Journal of Molecular Biology Volume: 420 Issue: 1-2 Pages: 40-55
  • DOI: 10.1016/j.jmb.2012.03.029
  • NAID: 120004186302
  • Peer Reviewed
• [Journal Article] A Three-Dimensional FRET Analysis to Construct an Atomic Model of the Actin-Tropomyosin Complex on a Reconstituted Thin Filament (2011)
  • Author(s): M. Miki, S. Makimura, T. Saitoh, M. Bunya, Y. Sugahara, Y. Ueno, C. Kimura-Sakiyama & H. Tobita
  • Journal Title: J. Mol. Biol Volume: 414 Pages: 765-782
  • NAID: 120003726650
  • Peer Reviewed
• [Journal Article] Interaction sites of tropomyosin in muscle thin filament as identified by site-directed spin-labeling (2011)
  • Author(s): K. Ueda, C. Kimura-Sakiyama, T. Aihara, M. Miki & T. Arata
  • Journal Title: Biophys. J. Volume: 100 Pages: 2432-2439
  • NAID: 120003726649
  • Peer Reviewed
• [Journal Article] Interaction Sites of Tropomyosin in Muscle Thin Filament as Identified by Site-Directed Spin-Labeling (2011)
  • Author(s): Ueda, K., Kimura-Sakiyama, C., Aihara, T., Miki, M., Arata, T.
  • Journal Title: Biophysical Journal Volume: 100 Issue: 10 Pages: 2432-2439
  • DOI: 10.1016/j.bpj.2011.03.021
  • NAID: 120003726649
  • Peer Reviewed
• [Journal Article] A Three-Dimensional FRET Analysis to Construct an Atomic Model of the Actin-Tropomyosin Complex on a Reconstituted Thin Filament (2011)
  • Author(s): Miki, M., Makimura, S., Saitoh, T., Bunya, M., Sugahara, Y., Ueno, Y., Kimura-Sakiyama, C., Tobita, H.
  • Journal Title: Journal of Molecular Biology Volume: 414 Issue: 5 Pages: 765-782
  • DOI: 10.1016/j.jmb.2011.10.033
  • NAID: 120003726650
  • Peer Reviewed
• [Journal Article] Switch action of Troponin on Muscle Thin Filament as Revealed by Spin Labeling and Pulsed EPR (2010)
  • Author(s): T. Aihara, M. Nakamura, S. Ueki, H. Hara, M. Miki, and T. Arata
  • Journal Title: J. Biol. Chem Volume: 285 Pages: 10671-10677
  • Peer Reviewed
• [Journal Article] Switch Action of Troponin on Muscle Thin Filament as Revealed by Spin Labeling and Pulsed ESR (2010)
  • Author(s): Aihara, Nakamura, Ueki, Hara, Mild, Arata
  • Journal Title: The Journal of Biological Chemistry Volume: 1285 Pages: 10671-1067
  • Peer Reviewed
• [Presentation] A Three-Dimensional FRET Analysis to Construct an Atomic Model of the Actin. Tropomyosin. Troponin Core Domain Complex on a Muscle Thin Filament (2012)
  • Author(s): Masao Miki
  • Organizer: International Workshop "from Structure to Dynamics : for Our Understanding of Protein-Protein Interactions"
  • Place of Presentation: 長浜
  • Year and Date: 2012-03-18
• [Presentation] A Three-Dimensional FRET Analysis to Locate the Troponin Core Domain on a Reconstituted Thin Filament (2012)
  • Author(s): 三木正雄、牧村敏史、菅原康之、山田隆太、文屋正志、飛田英孝
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 筑波大学
  • Year and Date: 2012-01-06
• [Presentation] A Three-Dimensional FRET Analysis to Locate the Troponin Core Domain on the Reconstituted Thin Filament (2012)
  • Author(s): 三木正雄、牧村敏史、菅原康之、山田隆太、文屋正志、斎藤隆弘、飛田英孝
  • Organizer: 生体運動合同研究班会議
  • Place of Presentation: 筑波大学
  • Year and Date: 2012-01-06
• [Presentation] 三次元FRET解析によるFアクチンートロポニンコアドメイン複合体のアトミックモデル構築 (2011)
  • Author(s): 三木正雄、牧村敏史、菅原康之、山田隆太、文屋正志、飛田英孝
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 兵庫県立大学姫路
  • Year and Date: 2011-09-18
• [Presentation] 三次元FRET解析によるFアクチンートロポニンコアドメイン複合体のアトミックモデル構築 (2011)
  • Author(s): 三木正雄、牧村敏史、菅原康之、山田隆太、文屋正志、飛田英孝
  • Organizer: 日本生物物理学会第49回年会
  • Place of Presentation: 兵庫県立大学姫路
  • Year and Date: 2011-09-08
• [Presentation] 三次元FRET解析によるFアクチンートロポミオシン複合体のアトミックモデル構築 (2011)
  • Author(s): 三木正雄,牧村敏史,文屋正志,斉藤隆弘,菅原康之,飛田英孝
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 大阪市大
  • Year and Date: 2011-01-07
• [Presentation] 三次元FRET解析によるFアクチン-トロポミオシン複合体のアトミックモデル構築 (2011)
  • Author(s): 三木正雄, 牧村敏史, 文屋正志, 斉藤隆弘, 菅原康之, 飛田英孝
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 大阪市立大学(大阪府)
  • Year and Date: 2011-01-07
• [Presentation] FRET between probes on Actin and Tropomyosin in the Reconstituted Thin Filament for Constructing an Atomic Model of the FA-Tm Complex (2010)
  • Author(s): 牧村敏史、文屋正志、斉藤隆弘、菅原康之、飛田英孝、三木正雄
  • Organizer: 日本生物物理学会第48回年会
  • Place of Presentation: 東北大学
  • Year and Date: 2010-09-20
• [Presentation] FRET between probes on Actin and Tropomyosin in the Reconstituted Thin Fila ment for Constructing an Atomic Model of the FA-Tm Complex (2010)
  • Author(s): 牧村敏史、文屋正志、斉藤隆弘、菅原康之、飛田英孝、三木正雄
  • Organizer: 日本生物理学会年会
  • Place of Presentation: 東北大学(宮城県)
  • Year and Date: 2010-09-20
• [Presentation] Three-Dimensional FRET Analysis for Constructing an Atomic Model of the Muscle Thin Filament (2010)
  • Author(s): 三木正雄、飛田英孝、斎藤隆広、牧村敏史、文屋正志
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 東京大学
  • Year and Date: 2010-01-10
• [Presentation] Three-Dimensional FRET Analysis for Constructing an Atomic Model of the Muscle Thin Filament (2010)
  • Author(s): 三木正雄, 飛田英孝, 斎藤隆広, 牧村敏史, 文屋正志
  • Organizer: 生体運動研究合同班会議
  • Place of Presentation: 生体運動研究合同班会議
  • Year and Date: 2010-01-10
• [Presentation] FRET between Residues on Actin and the N-terminal region(83-111) of Tropomyosin in the Reconstituted Thin Filament (2009)
  • Author(s): M. Bunya, S. Makimura, H. Tobita, and M. Miki
  • Organizer: 日本生物物理学会第47回年会
  • Place of Presentation: 徳島文理大学
  • Year and Date: 2009-11-01
• [Presentation] FRET between Residues on Actin and the N-terminal region (83-111) of Tropomyosin in the Reconstituted Thin Filament (2009)
  • Author(s): M.Bunya, S.Makimura, H.Tobita, M.Miki
  • Organizer: 日本生物物理学会第47回年会
  • Place of Presentation: 日本生物物理学会第47回年会
  • Year and Date: 2009-11-01