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Single-molecule tracking of myosins with genetically engineered amplifier domains

Nature Structural Biology volume 8, pages 226–229 (2001)Cite this article

Abstract

We combined protein engineering and single molecule measurements to directly record the step size of a series of myosin constructs with shortened and elongated artificial neck domains. Our results show that the step size has a clear linear dependence on the length of the neck domain and we also established that mechanical amplification in the myosin motor is based on a rotation of the neck domain relative to the actin-bound head. For all our constructs, including those with artificial necks, the magnitude of the neck rotation concurrent with the displacement step was ∼30°. The engineered change in the step size of myosin marks a significant advance in our ability to selectively modify the functional properties of molecular motors.

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Figure 1: Myosin molecules and step sizes.
Figure 2: Arrangement for recording single motor events.
Figure 3: Analysis of the step size behavior of myosin motors and constructs.

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Acknowledgements

We would like to acknowledge technical assistance from D.M. Hunt, U. Rühl, P. Uta and S. Zimmermann and help of the Zentrale Forschungswerkstätten with the construction of equipment. We thank M.L.W. Knetsch, S. Fujita-Becker and G. Tsiavaliaris for generous help. T.Q.P. Uyeda, S. Adlerstein, B. Sodeik, J. Kull and E. Ungewickell critically reviewed different versions of the manuscript and made many helpful suggestions. This work was supported by grants from the Deutsche Forschungsgemeinschaft (B.B., E.M.), Volkswagen-Stiftung (D.J.M., E.M.) and Max-Planck-Society (D.J.M.).

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Authors and Affiliations

  1. Molecular and Cellular Physiology, Medical School Hanover, Carl-Neuberg-Str. 1, Hannover, 30625, Germany

    Christine Ruff, Bernhard Brenner & Edgar Meyhöfer

  2. Department of Biophysics, Max-Planck-Institute for Medical Research, Jahnstr. 29, Heidelberg, 69120, Germany

    Marcus Furch & Dietmar J. Manstein

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  1. Christine Ruff
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  2. Marcus Furch
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Correspondence to Edgar Meyhöfer.

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Ruff, C., Furch, M., Brenner, B. et al. Single-molecule tracking of myosins with genetically engineered amplifier domains. Nat Struct Mol Biol 8, 226–229 (2001). https://doi.org/10.1038/84962

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