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Regulation by Ca2+-calmodulin of the actin-bundling activity ofPhysarum 210-kDa protein

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Summary

From the plasmodia of a lower eukaryote,Physarum polycephalum, we have previously purified a 210-kDa protein that showed similar properties to those of smooth muscle caldesmon. Further characterization of the 210-kDa protein revealed that it bundled actin filaments. This bundling activity was inhibited by calmodulin in the presence of Ca2+. Unlike smooth muscle caldesmon, the 210-kDa protein bundled actin filaments whether or not a reducing agent, such as dithiothreitol, was present. The protein was shown to have two (or more) different actin-binding sites which were classified into salt-sensitive and salt-insensitive sites. Electron microscopy revealed that the 210-kDa protein was an elongated molecule (mean length, 97 ± 25 nm) which was bent in the middle. The Stokes radius and sedimentation coefficient of the 210-kDa protein were 130 Å and 2.9 S, respectively. An immunofluorescence study revealed that the 210-kDa protein colocalized with the bundles of actin filaments in thin-spread preparations ofPhysarum plasmodia, suggesting that the 210-kDa protein was regulating the appearance and disappearance of the actin bundles that are associated with the contraction-relaxation cycle of the plasmodia.

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Authors and Affiliations

  1. Department of Pharmacology, Gunma University School of Medicine, Maebashi, 371, Gunma, Japan

    Ryoki Ishikawa, Tsuyoshi Okagaki & Kazuhiro Kohama

  2. Department of Cell Biology and Anatomy, Cornell University Medical College, 10021, New York, NY, USA

    Tsuyoshi Okagaki

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  1. Ryoki Ishikawa
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  2. Tsuyoshi Okagaki
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  3. Kazuhiro Kohama
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Ishikawa, R., Okagaki, T. & Kohama, K. Regulation by Ca2+-calmodulin of the actin-bundling activity ofPhysarum 210-kDa protein. J Muscle Res Cell Motil 13, 321–328 (1992). https://doi.org/10.1007/BF01766460

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  • DOI: https://doi.org/10.1007/BF01766460

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