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An actin-dependent conformational change in myosin

Nature Structural Biology volume 10, pages 402408 (2003) | Download Citation

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Abstract

Conformational changes within myosin lead to its movement relative to an actin filament. Several crystal structures exist for myosin bound to various nucleotides, but none with bound actin. Therefore, the effect of actin on the structure of myosin is poorly understood. Here we show that the swing of smooth muscle myosin lever arm requires both ADP and actin. This is the first direct observation that a conformation of myosin is dependent on actin. Conformational changes within myosin were monitored using fluorescence resonance energy transfer techniques. A cysteine-reactive probe is site-specifically labeled on a 'cysteine-light' myosin variant, in which the native reactive cysteines were removed and a cysteine engineered at a desired position. Using this construct, we show that the actin-dependent ADP swing causes an 18 Å change in distance between a probe on the 25/50 kDa loop on the catalytic domain and a probe on the regulatory light chain, corresponding to a 23° swing of the light-chain domain.

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Acknowledgements

This work was supported by NIH grants to P.R.S. and H.L.S.

Author information

Author notes

    • Ming Xiao

    Current address: Cardiovascular Research Institute, University of California, San Francisco, California 94143, USA.

Affiliations

  1. Department of Physics, University of Illinois, 1110 West Green Street, Urbana, Illinois 61801, USA.

    • Ming Xiao
    • , Jeff G. Reifenberger
    • , Pinghua Ge
    •  & Paul R. Selvin
  2. Department of Physiology, University of Pennsylvania School of Medicine, 3700 Hamilton Walk, Philadelphia, Pennsylvania 19104-6085, USA.

    • Amber L. Wells
    • , Corry Baldacchino
    • , Li-Qiong Chen
    •  & H. Lee Sweeney
  3. Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61801, USA.

    • Paul R. Selvin

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Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to H. Lee Sweeney or Paul R. Selvin.

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DOI

https://doi.org/10.1038/nsb916

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