Use of thin filament reconstituted muscle fibres to probe the mechanism of force generation

  • Masataka KawaiEmail author
  • Shin’ichi Ishiwata
Original Paper


The technique of selective removal of the thin filament by gelsolin in bovine cardiac muscle fibres, and reconstitution of the thin filament from isolated proteins is reviewed, and papers that used reconstituted preparations are discussed. By comparing the results obtained in the absence/presence of regulatory proteins tropomyosin (Tm) and troponin (Tn), it is concluded that the role of Tm and Tn in force generation is not only to expose the binding site of actin to myosin, but also to modify actin for better stereospecific and hydrophobic interaction with myosin. This conclusion is further supported by experiments that used a truncated Tm mutant and the temperature study of reconstituted fibres. The conclusion is consistent with the hypothesis that there are three states in the thin filament: blocked state, closed state, and open state. Tm is the major player to produce these effects, with Tn playing the role of Ca2+ sensing and signal transmission mechanism. Experiments that changed the number of negative charges at the N-terminal finger of actin demonstrates that this part of actin is essential to promote the strong interaction between actin and myosin molecules, in addition to the well-known weak interaction that positions the myosin head at the active site of actin prior to force generation.


Actin Regulatory proteins Tropomyosin Troponin Myosin Gelsolin Cross-bridge kinetics Sinusoidal analysis BDM 


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We would like to thank Dr. Hideaki Fujita for drawing Fig. 1, and to Dr. Madoka Suzuki and Ms. Kristen Stanton for critical reading of the manuscript. This work was supported in part by an NIH grant HL70041 to MK, and by Grants-in-Aid for Specially Promoted Research and for the 21st Century COE program (Physics of Self-organization Systems), and by “Establishment of Consolidated Research Institute for Advanced Science and Medical Care” from the Ministry of Education, Sports, Culture, Science and Technology of Japan to SI. The contents of this work are solely the responsibility of the authors and do not necessarily represent the official view of awarding organizations.


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© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  1. 1.Department of Anatomy and Cell Biology, College of MedicineThe University of IowaIowa CityUSA
  2. 2.Department of Physics, School of Science and Engineering, and Advanced Research Institute for Science and EngineeringWaseda UniversityTokyoJapan

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