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Filamin and gelsolin influence Ca2+-sensitivity of smooth muscle thin filaments

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Summary

Sheep aorta thin filaments were prepared by ultracentrifugation of an ATP-containing extract in the presence of different concentrations of ethanediol. Thin filaments prepared without ethanediol contained small quantities of tropomyosin (0.027 Tm:actin) and caldesmon (0.017 CD:actin) and activated the MgATPase of skeletal myosin independently of Ca2+. Ultracentrifugation in the presence of 10–20% ethanediol resulted in preparation of thin filaments with increased content of tropomyosin (0.17 Tm:actin) and caldesmon (0.04 CD:actin). These thin filaments possessed high Ca2+-sensitivity in activation of skeletal muscle myosin ATPase. Besides actin, tropomyosin and caldesmon, thin filaments contained gelsolin and filamin. Gelsolin content (0.007 gelsolin:actin) was independent of the presence of ethanediol. The filamin content decreased from 0.015 to 0.007 mol:mol actin when the ethanediol concentration was increased from 0 to 20%, and was negatively correlated with the Ca2+ sensitivity of thin filaments. In a reconstituted system, pure filamin or gelsolin affected caldesmon regulation of actomyosin ATPase. Gelsolin (0.01:actin) reduced the inhibition of actomyosin ATPase caused by caldesmon and increased the potency of Ca2+-calmodulin in reversing this inhibition. Filamin (0.007:actin) also decreased the inhibitory action of caldesmon on actin-activated myosin ATPase and also potentiated the reversal of this inhibition by calmodulin. We conclude that minor components of smooth muscle thin filaments (gelsolin and filamin) significantly modify caldesmon mediated regulation of actomyosin ATPase. We suggest a tropomyosin-mediated mechanism by which filamin or gelsolin could exert similar effects.

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Authors and Affiliations

  1. Department of Biochemistry, School of Biology, Moscow State University, 119889, Moscow, Russia

    Nikolai B. Gusev

  2. Department of Cardiac Medicine, National Heart and Lung Institute, Dovehouse Street, SW3 6LY, London, UK

    Kevin Pritchard, Julie L. Hodgkinson & Steven B. Marston

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  1. Nikolai B. Gusev
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  2. Kevin Pritchard
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  3. Julie L. Hodgkinson
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Gusev, N.B., Pritchard, K., Hodgkinson, J.L. et al. Filamin and gelsolin influence Ca2+-sensitivity of smooth muscle thin filaments. J Muscle Res Cell Motil 15, 672–681 (1994). https://doi.org/10.1007/BF00121074

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