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Load-dependent kinetics of force production by smooth muscle myosin measured with optical tweezers

Nature Cell Biology volume 5, pages 980–986 (2003)Cite this article

Abstract

Muscle contraction is driven by the cyclical interaction of myosin with actin, coupled with ATP hydrolysis. Myosin attaches to actin, forming a crossbridge that produces force and movement as it tilts1 or rocks into subsequent bound states2 before finally detaching. It has been hypothesized that the kinetics of one or more of these mechanical transitions are dependent on load, allowing muscle to shorten quickly under low load, but to sustain tension economically, with slowly cycling crossbridges under high load conditions2,3,4,5,6. The idea that muscle biochemistry depends on mechanical output is termed the 'Fenn effect'. However, the molecular details of how load affects the kinetics of a single crossbridge are unknown. Here, we describe a new technique based on optical tweezers to rapidly apply force to a single smooth muscle myosin crossbridge. The crossbridge produced movement in two phases that contribute 4 nm + 2 nm of displacement. Duration of the first phase depended in an exponential manner on the amplitude of applied load. Duration of the second phase was much less affected by load, but was significantly shorter at high ATP concentration. The effect of load on the lifetime of the bound crossbridge is to prolong binding when load is high, but to accelerate release when load is low or negative.

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Figure 1: Effect of load on single-molecule mechanical interactions measured for SmS1.
Figure 2: (a, b) Ensemble-averaged attachment events at two ATP concentrations.
Figure 3: Ensemble-averaged attachment events in the absence or presence of ATP.
Figure 4: Effect of force on the rate constant k1.

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Acknowledgements

We would like to thank The Royal Society, the Medical Research Council and the Biotechnology and Biological Sciences Research Council for financial support, A.F. Huxley, D.R. Trentham, P.M. Bayley, and D.C.S. White for helpful discussions and comments, and M.L. Bartoo and Y. E. Goldman for help and ideas in the early stages of this work.

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Authors and Affiliations

  1. Division of Physical Biochemistry, NIMR, The Ridgeway Mill Hill, London, NW7 1AA, UK

    Claudia Veigel, Justin E. Molloy & Stephan Schmitz

  2. MRC, Laboratory for Molecular Biology, Cambridge, CB2 2QH, UK

    John Kendrick-Jones

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  1. Claudia Veigel
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Correspondence to Claudia Veigel.

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Veigel, C., Molloy, J., Schmitz, S. et al. Load-dependent kinetics of force production by smooth muscle myosin measured with optical tweezers. Nat Cell Biol 5, 980–986 (2003). https://doi.org/10.1038/ncb1060

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