Mechanical coordination in motor ensembles revealed using engineered artificial myosin filaments


The sarcomere of muscle is composed of tens of thousands of myosin motors that self-assemble into thick filaments and interact with surrounding actin-based thin filaments in a dense, near-crystalline hexagonal lattice1. Together, these actin–myosin interactions enable large-scale movement and force generation, two primary attributes of muscle. Research on isolated fibres has provided considerable insight into the collective properties of muscle, but how actin–myosin interactions are coordinated in an ensemble remains poorly understood2. Here, we show that artificial myosin filaments, engineered using a DNA nanotube scaffold, provide precise control over motor number, type and spacing. Using both dimeric myosin V- and myosin VI-labelled nanotubes, we find that neither myosin density nor spacing has a significant effect on the gliding speed of actin filaments. This observation supports a simple model of myosin ensembles as energy reservoirs that buffer individual stochastic events to bring about smooth, continuous motion. Furthermore, gliding speed increases with cross-bridge compliance, but is limited by Brownian effects. As a first step to reconstituting muscle motility, we demonstrate human β-cardiac myosin-driven gliding of actin filaments on DNA nanotubes.

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Figure 1: Formation of synthetic myosin filaments using DNA nanotubes.
Figure 2: Density and number of myosins on synthetic filaments have negligible effect on gliding speeds.
Figure 3: Stochastic simulation of actin gliding along myosin filaments predicts actin length-independent gliding speed for sufficiently high motor number.
Figure 4: Actin gliding on synthetic human β-cardiac myosin filaments.


  1. 1

    Huxley, H. E. The mechanism of muscular contraction. Science 164, 1356–1365 (1969).

  2. 2

    Guérin, T., Prost, J., Martin, P. & Joanny, J. F. Coordination and collective properties of molecular motors: theory. Curr. Opin. Cell Biol. 22, 14–20 (2010).

  3. 3

    De La Cruz, E. M. & Ostap, E. M. Relating biochemistry and function in the myosin superfamily. Curr. Opin. Cell Biol. 16, 61–67 (2004).

  4. 4

    Uyeda, T. Q., Kron, S. J. & Spudich, J. A. Myosin step size. Estimation from slow sliding movement of actin over low densities of heavy meromyosin. J. Mol. Biol. 214, 699–710 (1990).

  5. 5

    Sommese, R. F. et al. Molecular consequences of the R453C hypertrophic cardiomyopathy mutation on human β-cardiac myosin motor function. Proc. Natl Acad. Sci. USA 110, 12607–12612 (2013).

  6. 6

    Harris, D. E. & Warshaw, D. M. Smooth and skeletal muscle myosin both exhibit low duty cycles at zero load in vitro. J. Biol. Chem. 268, 14764–14768 (1993).

  7. 7

    Piazzesi, G. et al. Skeletal muscle performance determined by modulation of number of myosin motors rather than motor force or stroke size. Cell 131, 784–795 (2007).

  8. 8

    Spudich, J. A. Hypertrophic and dilated cardiomyopathy: four decades of basic research on muscle lead to potential therapeutic approaches to these devastating genetic diseases. Biophys. J. 106, 1236–1249 (2014).

  9. 9

    Baker, J. E., Brosseau, C., Joel, P. B. & Warshaw, D. M. The biochemical kinetics underlying actin movement generated by one and many skeletal muscle myosin molecules. Biophys. J. 82, 2134–2147 (2002).

  10. 10

    Derr, N. D. et al. Tug-of-war in motor protein ensembles revealed with a programmable DNA origami scaffold. Science 338, 662–665 (2012).

  11. 11

    Hariadi, R. F., Cale, M. & Sivaramakrishnan, S. Myosin lever arm directs collective motion on cellular actin network. Proc. Natl Acad. Sci. USA 111, 4091–4096 (2014).

  12. 12

    Walcott, S., Warshaw, D. M. & Debold, E. P. Mechanical coupling between myosin molecules causes differences between ensemble and single-molecule measurements. Biophys. J. 103, 501–510 (2012).

  13. 13

    Rock, R. S., Rief, M., Mehta, A. D. & Spudich, J. A. In vitro assays of processive myosin motors. Methods 22, 373–381 (2000).

  14. 14

    Lu, H. et al. Collective dynamics of elastically coupled myosin V motors. J. Biol. Chem. 287, 27753–27761 (2012).

  15. 15

    Yin, P. et al. Programming DNA tube circumferences. Science 321, 824–826 (2008).

  16. 16

    Ali, M. Y. et al. Myosin Va and myosin VI coordinate their steps while engaged in an in vitro tug of war during cargo transport. Proc. Natl Acad. Sci. USA 108, E535–E541 (2011).

  17. 17

    Hilbert, L., Cumarasamy, S., Zitouni, N. B., Mackey, M. C. & Lauzon, A. M. The kinetics of mechanically coupled myosins exhibit group size-dependent regimes. Biophys. J. 105, 1466–1474 (2013).

  18. 18

    Rief, M. et al. Myosin-V stepping kinetics: a molecular model for processivity. Proc. Natl Acad. Sci. USA 97, 9482–9486 (2000).

  19. 19

    Rock, R. S. et al. Myosin VI is a processive motor with a large step size. Proc. Natl Acad. Sci. USA 98, 13655–13659 (2001).

  20. 20

    Gebhardt, J. C. M., Clemen, A. E. M., Jaud, J. & Rief, M. Myosin-V is a mechanical ratchet. Proc. Natl Acad. Sci. USA 103, 8680–8685 (2006).

  21. 21

    Veigel, C., Molloy, J. E., Schmitz, S. & Kendrick-Jones, J. Load-dependent kinetics of force production by smooth muscle myosin measured with optical tweezers. Nature Cell Biol. 5, 980–986 (2003).

  22. 22

    Walsh, R., Rutland, C., Thomas, R. & Loughna, S. Cardiomyopathy: a systematic review of disease-causing mutations in myosin heavy chain 7 and their phenotypic manifestations. Cardiology 115, 49–60 (2010).

  23. 23

    Weith, A. et al. Unique single molecule binding of cardiac myosin binding protein-C to actin and phosphorylation-dependent inhibition of actomyosin motility requires 17 amino acids of the motif domain. J. Mol. Cell. Cardiol. 52, 219–227 (2012).

  24. 24

    Resnicow, D. I., Deacon, J. C., Warrick, H. M., Spudich, J. A. & Leinwand, L. A. Functional diversity among a family of human skeletal muscle myosin motors. Proc. Natl Acad. Sci. USA 107, 1053–1058 (2010).

  25. 25

    Trybus, K. M., Freyzon, Y., Faust, L. Z. & Sweeney, H. L. Spare the rod, spoil the regulation: the necessity for a myosin rod. Proc. Natl Acad. Sci. USA 94, 48–52 (1997).

  26. 26

    Huang, J., Nagy, S. S., Koide, A., Rock, R. S. & Koide, S. A peptide tag system for facile purification and single-molecule immobilization. Biochemistry 48, 11834–11836 (2009).

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The authors thank M. Westfall, D. Smith and L. Hilbert for useful discussions. Research was funded by the American Heart Association Scientist Development Grant (13SDG14270009), National Institutes of Health (NIH) grants 1DP2 CA186752-01 and 1-R01-GM-105646-01-A1 to S.S. and NIH grants GM33289 and HL117138 to J.A.S. R.F.S. is a Life Sciences Research Foundation Fellow. R.E.T. is supported by the NIH (F32 HL123247-02) and A.S.A. is supported by a Lucile Packard CHRI Postdoctoral Award.

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R.F.H., R.F.S., A.S.A., R.E.T., J.A.S. and S.S. planned and designed experiments. R.F.H., R.F.S., A.S.A., R.E.T. and S.Su. performed experiments and analysed the results. R.F.H. and S.S. performed the mathematical modelling. R.F.H., R.F.S., J.A.S. and S.S. wrote the manuscript.

Correspondence to S. Sivaramakrishnan.

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Hariadi, R., Sommese, R., Adhikari, A. et al. Mechanical coordination in motor ensembles revealed using engineered artificial myosin filaments. Nature Nanotech 10, 696–700 (2015).

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