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

Masataka Kawai, Shin'ichi Ishiwata

    Research output: Contribution to journalArticlepeer-review

    22 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish
    Pages (from-to)455-468
    Number of pages14
    JournalJournal of Muscle Research and Cell Motility
    Volume27
    Issue number5-7
    DOIs
    Publication statusPublished - 2006 May

    Keywords

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

    ASJC Scopus subject areas

    • Physiology
    • Clinical Biochemistry
    • Endocrinology
    • Cell Biology

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