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Single-molecule stepping and structural dynamics of myosin X

Nature Structural & Molecular Biology volume 17, pages 485–491 (2010)Cite this article

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Abstract

Myosin X is an unconventional myosin with puzzling motility properties. We studied the motility of dimerized myosin X using the single-molecule fluorescence techniques polTIRF, FIONA and Parallax to measure the rotation angles and three-dimensional position of the molecule during its walk. It was found that Myosin X steps processively in a hand-over-hand manner following a left-handed helical path along both single actin filaments and bundles. Its step size and velocity are smaller on actin bundles than individual filaments, suggesting myosin X often steps onto neighboring filaments in a bundle. The data suggest that a previously postulated single α-helical domain mechanically extends the lever arm, which has three IQ motifs, and either the neck-tail hinge or the tail is flexible. These structural features, in conjunction with the membrane- and microtubule-binding domains, enable myosin X to perform multiple functions on varied actin structures in cells.

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Figure 1: Processivity and stepping rates of labeled myosin X (MyoX-LA-QD655) actin filaments and actin bundles.
Figure 2: polTIRF results for MyoX-BRCaM stepping on an actin filament immobilized on the glass surface.
Figure 3: Alternating step size supports the hand-over-hand model of myosin X.
Figure 4: MyoX-CT-QD655 shows different stepping behavior along actin filaments and bundles.
Figure 5: Parallax tracking of myosin X molecules stepping along suspended actin filaments and bundles at 1 μM Mg-ATP.
Figure 6: Radial amplitudes of helical paths of myosin X molecules stepping along suspended actins.

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Acknowledgements

We acknowledge discussion and comments by J.H. Lewis and J.F. Beausang. We thank T.M. Svitkina and C. Yang for characterization of fascin-bundled actin by electron microscopy (US National Institutes of Health (NIH) grant RR 22482) and K. Homma (University of Massachusetts Medical School) for the GFP–Myosin X HMM construct. Bifunctional rhodamine (BR-I2) was a generous gift from J.E.T. Corrie (UK Medical Research Council National Institute for Medical Research). This work was supported by US National Science Foundation Nanoscale Science and Engineering Center grant DMR-0425780 through the Nano/Bio Interface Center and NIH grant GM086352.

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Authors and Affiliations

  1. Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Yujie Sun, Mark E Arsenault & Yale E Goldman

  2. University of Massachusetts Medical School, North Worcester, Massachusetts, USA

    Osamu Sato & Mitsuo Ikebe

  3. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Felix Ruhnow

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Correspondence to Yale E Goldman.

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Sun, Y., Sato, O., Ruhnow, F. et al. Single-molecule stepping and structural dynamics of myosin X. Nat Struct Mol Biol 17, 485–491 (2010). https://doi.org/10.1038/nsmb.1785

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