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Evidence for cleft closure in actomyosin upon ADP release

Nature Structural Biology volume 7, pages 1147–1155 (2000)Cite this article

Abstract

Structural insights into the interaction of smooth muscle myosin with actin have been provided by computer-based fitting of crystal structures into three-dimensional reconstructions obtained by electron cryomicroscopy, and by mapping of structural and dynamic changes in the actomyosin complex. The actomyosin structures determined in the presence and absence of MgADP differ significantly from each other, and from all crystallographic structures of unbound myosin. Coupled to a complex movement (∼34 Å) of the light chain binding domain upon MgADP release, we observed a ∼9° rotation of the myosin motor domain relative to the actin filament, and a closure of the cleft that divides the actin binding region of the myosin head. Cleft closure is achieved by a movement of the upper 50 kDa region, while parts of the lower 50 kDa region are stabilized through strong interactions with actin. This model supports a mechanism in which binding of MgATP at the active site opens the cleft and disrupts the interface, thereby releasing myosin from actin.

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Figure 1: Stereo views of the final smooth muscle myosin models and the corresponding 3D reconstructions of actomyosin a, in the presence of MgADP and b, in the absence of nucleotide.
Figure 2: Best fits for crystal structures to the 3D rigor reconstructions.
Figure 3: Depiction of the light chain domain movement.
Figure 4: Differences between the final models and the corresponding reconstructions (discrepancy maps).
Figure 5: Depiction of conformational changes and structural flexibility in the MgADP state and the rigor state.

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Acknowledgements

This work was supported by National Institutes of Health grants to D.J.D., S.L. and K.M.T. and a National Science Foundation grant to S.L. for the support of G.O. The authors acknowledge funds from the W.M. Keck Foundation.

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Authors and Affiliations

  1. The Burnham Institute, La Jolla, 92037, California, USA

    Niels Volkmann & Dorit Hanein

  2. The Rosenstiel Basic Medical Sciences Research Center and The W.M. Keck Institute for Cellular Visualization, Brandeis University, Waltham, 02254, Massachusetts, USA

    Greta Ouyang & David J. DeRosier

  3. Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, 05405, Vermont, USA

    Kathleen M. Trybus & Susan Lowey

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Correspondence to Dorit Hanein.

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Volkmann, N., Hanein, D., Ouyang, G. et al. Evidence for cleft closure in actomyosin upon ADP release. Nat Struct Mol Biol 7, 1147–1155 (2000). https://doi.org/10.1038/82008

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