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Direct observation of the myosin-Va power stroke and its reversal

Nature Structural & Molecular Biology volume 17, pages 590–595 (2010)Cite this article

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Abstract

Complex forms of cellular motility, including cell division, organelle trafficking or signal amplification in the auditory system, require strong coordination of the myosin motors involved. The most basic mechanism of coordination is via direct mechanical interactions of individual motor heads leading to modification of their mechanochemical cycles. Here we used an optical trap–based assay to investigate the reversibility of the force-generating conformational change (power stroke) of single myosin-Va motor heads. By applying load to the head shortly after binding to actin, we found that, at a certain load, the power stroke could be reversed, and the head fluctuated between an actin-bound pre– and a post–power stroke conformation. This load-dependent mechanical instability might be critical to coordinate the heads of processive, dimeric myosin-Va. Nonlinear response to load leading to coordination or oscillations amongst motors might be relevant for many cellular functions.

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Figure 1: Single MVS1 molecules interacting with actin under load.
Figure 2: Statistical analysis of reversals and recoveries.
Figure 3: Kinetics of pre- and post-states.
Figure 4: Effect of load on a single MVS1 motor head.

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Acknowledgements

We thank C. Schmidt and J. Molloy for stimulating discussions, E. Harvey for technical assistance and J. Hammer III (Laboratories of Cell Biology, National Heart, Lung, and Blood Institute, US National Institutes of Health) for kindly supplying the myosin-Va clone. We are also grateful to the UK Medical Research Council, The Royal Society UK and the US National Institutes of Health for grant support.

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Authors and Affiliations

  1. Laboratory of Molecular Physiology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA

    James R Sellers

  2. Physical Biochemistry, National Institute for Medical Research, London, UK

    Claudia Veigel

  3. Department of Cellular Physiology, Ludwig Maximilians University Munich, Munich, Germany

    Claudia Veigel

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  1. James R Sellers
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  2. Claudia Veigel
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Contributions

J.R.S. expressed and purified MVS1; C.V. carried out experiments and data analysis; J.R.S. and C.V. wrote the paper.

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Correspondence to Claudia Veigel.

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The authors declare no competing financial interests.

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Supplementary Figures 1–5, Supplementary Table 1 and Supplementary Discussion (PDF 8752 kb)

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Sellers, J., Veigel, C. Direct observation of the myosin-Va power stroke and its reversal. Nat Struct Mol Biol 17, 590–595 (2010). https://doi.org/10.1038/nsmb.1820

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