Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments

Masataka Kawai, Takanori Kido, Martin Vogel, Rainer H A Fink, Shin'ichi Ishiwata

    Research output: Contribution to journalArticle

    27 Citations (Scopus)

    Abstract

    The temperature dependence of sliding velocity, force and the number of cross-bridges was studied on regulated actin filaments (reconstituted thin filaments) when they were placed on heavy meromyosin (HMM) attached to a glass surface. The regulated actin filaments were used because our previous study on muscle fibres demonstrated that the temperature effect was much reduced in the absence of regulatory proteins. A fluorescently labelled thin filament was attached to the gelsolin-coated surface of a polystyrene bead. The bead was trapped by optical tweezers, and HMM-thin filament interaction was performed at 20-35°C to study the temperature dependence of force at the single-molecule level. Our experiments showed that there was a small increase in force with temperature (Q10 =1.43) and sliding velocity (Q10 =1.46). The small increase in force was correlated with the small increase in the number of cross-bridges (Q10 =1.49), and when force was divided by the number of cross-bridges, the result did not depend on the temperature (Q10 =1.03). These results demonstrate that the force each cross-bridge generates is fixed and independent of temperature. Our additional experiments demonstrate that tropomyosin (Tm) in the presence of troponin (Tn) and Ca2+ enhances both force and velocity, and a truncated mutant, Δ23Tm, diminishes force and velocity. These results are consistent with the hypothesis that Tm in the presence of Tn and Ca2+ exerts a positive allosteric effect on actin to make actomyosin linkage more secure so that larger forces can be generated.

    Original languageEnglish
    Pages (from-to)877-887
    Number of pages11
    JournalJournal of Physiology
    Volume574
    Issue number3
    DOIs
    Publication statusPublished - 2006 Aug

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    Myosin Subfragments
    Actin Cytoskeleton
    Temperature
    Tropomyosin
    Troponin
    Optical Tweezers
    Gelsolin
    Actomyosin
    Polystyrenes
    Glass
    Actins
    Muscles
    Proteins

    ASJC Scopus subject areas

    • Physiology

    Cite this

    Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments. / Kawai, Masataka; Kido, Takanori; Vogel, Martin; Fink, Rainer H A; Ishiwata, Shin'ichi.

    In: Journal of Physiology, Vol. 574, No. 3, 08.2006, p. 877-887.

    Research output: Contribution to journalArticle

    Kawai, Masataka ; Kido, Takanori ; Vogel, Martin ; Fink, Rainer H A ; Ishiwata, Shin'ichi. / Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments. In: Journal of Physiology. 2006 ; Vol. 574, No. 3. pp. 877-887.
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