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Engineering controllable bidirectional molecular motors based on myosin

Nature Nanotechnology volume 7, pages 252–256 (2012)Cite this article

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Abstract

Cytoskeletal motors drive the transport of organelles and molecular cargoes within cells1 and have potential applications in molecular detection and diagnostic devices2,3. Engineering molecular motors with controllable properties will allow selective perturbation of mechanical processes in living cells and provide optimized device components for tasks such as molecular sorting and directed assembly3. Biological motors have previously been modified by introducing activation/deactivation switches that respond to metal ions4,5 and other signals6. Here, we show that myosin motors can be engineered to reversibly change their direction of motion in response to a calcium signal. Building on previous protein engineering studies7,8,9,10,11 and guided by a structural model12 for the redirected power stroke of myosin VI, we have constructed bidirectional myosins through the rigid recombination of structural modules. The performance of the motors was confirmed using gliding filament assays and single fluorophore tracking. Our strategy, in which external signals trigger changes in the geometry and mechanics of myosin lever arms, should make it possible to achieve spatiotemporal control over a range of motor properties including processivity, stride size13 and branchpoint turning14.

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Figure 1: Engineered myosin designs.
Figure 2: Gliding filament assays of engineered myosins.
Figure 3: Processive motility of a dimeric controllable motor.

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Acknowledgements

The authors thank M.W. Elting for valuable discussions and assistance, R. Cooke for providing full-length muscle myosin, and S. Sutton for providing purified actin. This work was supported by a Pew Scholars Award (to Z.B.), the NIH (grant no. DP2 OD004690 to Z.B.), an NSF Graduate Research Fellowship (to L.C.), an AHA Predoctoral Fellowship (to M.N.) and a Stanford Graduate Fellowship (to T.D.S.).

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Authors and Affiliations

  1. Department of Bioengineering, Stanford University, Stanford, 94305, California, USA

    Lu Chen, Muneaki Nakamura, Tony D. Schindler, David Parker & Zev Bryant

  2. Department of Chemistry, Stanford University, Stanford, 94305, California, USA

    Muneaki Nakamura

  3. Department of Structural Biology, Stanford University School of Medicine, Stanford, 94305, California, USA

    Zev Bryant

Authors
  1. Lu Chen
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Contributions

L.C., M.N. and Z.B. designed the molecular motor constructs. L.C., M.N. and T.D.S. performed experiments on fixed directionality motors. L.C. and M.N. performed experiments on bidirectional motors. L.C. analysed the experimental data. D.P., L.C. and M.N. performed structural modelling. Z.B. conceived and supervised the project. L.C. and Z.B. wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Zev Bryant.

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The authors declare no competing financial interests.

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Chen, L., Nakamura, M., Schindler, T. et al. Engineering controllable bidirectional molecular motors based on myosin. Nature Nanotech 7, 252–256 (2012). https://doi.org/10.1038/nnano.2012.19

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