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Remote control of myosin and kinesin motors using light-activated gearshifting

Nature Nanotechnology volume 9, pages 693–697 (2014)Cite this article

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Abstract

Cytoskeletal motors perform critical force generation and transport functions in eukaryotic cells1,2. Engineered modifications of motor function provide direct tests of protein structure–function relationships and potential tools for controlling cellular processes or for harnessing molecular transport in artificial systems3,4. Here, we report the design and characterization of a panel of cytoskeletal motors that reversibly change gears—speed up, slow down or switch directions—when exposed to blue light. Our genetically encoded structural designs incorporate a photoactive protein domain to enable light-dependent conformational changes in an engineered lever arm. Using in vitro motility assays, we demonstrate robust spatiotemporal control over motor function and characterize the kinetics of the optical gearshifting mechanism. We have used a modular approach to create optical gearshifting motors for both actin-based and microtubule-based transport.

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Figure 1: Molecular design, proposed switching mechanism and in vitro performance of MyLOV constructs.
Figure 2: Dynamic control of speed in MyLOVChar.
Figure 3: Design, structure and in vitro performance of engineered kinesin-14 motors.

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Acknowledgements

The authors thank J-C. Liao, T. Omabegho, D.J. Cipriano, P.V. Ruijgrok and M.W. Elting for technical assistance, S. Sutton and H.M. Warrick for providing purified actin, and M.J. Footer for providing a gelsolin expression plasmid. This work was supported by a Pew Scholars Award, National Institutes of Health grants DP2 OD004690 (to Z.B.) and P01GM051487 (to E.N.), an AHA Predoctoral Fellowship (to M.N.), a National Science Foundation Graduate Research Fellowship (to L.C.) and a Stanford Graduate Fellowship (to T.D.S.). E.N. is a Howard Hughes Medical Institute investigator.

Author information

Authors and Affiliations

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

    Muneaki Nakamura

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

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

  3. Biophysics Graduate Group, University of California, Berkeley, 94720, California, USA

    Stuart C. Howes

  4. Department of Molecular and Cell Biology, University of California, Berkeley, 94720, California, USA

    Eva Nogales

  5. Howard Hughes Medical Institute, University of California, Berkeley, 94720, California, USA

    Eva Nogales

  6. Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, 94720, California, USA

    Eva Nogales

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

    Zev Bryant

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  1. Muneaki Nakamura
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Contributions

M.N. designed myosin constructs and performed myosin experiments. T.D.S. assisted with myosin research. L.C. and M.N. designed and assayed Ncd constructs, analysed all motility data and provided samples and assistance for electron microscopy. S.C.H. performed cryoelectron microscopy and obtained three-dimensional reconstructions. E.N. supervised cryoelectron microscopy research. Z.B. conceived and supervised the overall project. M.N., Z.B., L.C. and S.C.H. contributed to writing 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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Nakamura, M., Chen, L., Howes, S. et al. Remote control of myosin and kinesin motors using light-activated gearshifting. Nature Nanotech 9, 693–697 (2014). https://doi.org/10.1038/nnano.2014.147

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