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Mechanical Characterization of One-Headed Myosin-V Using Optical Tweezers

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  • Tomonobu M Watanabe
  • Atsuko H Iwane
  • Hiroto Tanaka
  • Mitsuo Ikebe
  • Toshio Yanagida

Abstract

Class V myosin (myosin-V) is a cargo transporter that moves along an actin filament with large (∼36-nm) successive steps. It consists of two heads that each includes a motor domain and a long (23 nm) neck domain. One of the more popular models describing these steps, the hand-over-hand model, assumes the two-headed structure is imperative. However, we previously succeeded in observing successive large steps by one-headed myosin-V upon optimizing the angle of the acto-myosin interaction. In addition, it was reported that wild type myosin-VI and myosin-IX, both one-headed myosins, can also generate successive large steps. Here, we describe the mechanical properties (stepsize and stepping kinetics) of successive large steps by one-headed and two-headed myosin-Vs. This study shows that the stepsize and stepping kinetics of one-headed myosin-V are very similar to those of the two-headed one. However, there was a difference with regards to stability against load and the number of multisteps. One-headed myosin-V also showed unidirectional movement that like two-headed myosin-V required 3.5 kBT from ATP hydrolysis. This value is also similar to that of smooth muscle myosin-II, a non-processive motor, suggesting the myosin family uses a common mechanism for stepping regardless of the steps being processive or non-processive. In this present paper, we conclude that one-headed myosin-V can produce successive large steps without following the hand-over-hand mechanism.

Suggested Citation

  • Tomonobu M Watanabe & Atsuko H Iwane & Hiroto Tanaka & Mitsuo Ikebe & Toshio Yanagida, 2010. "Mechanical Characterization of One-Headed Myosin-V Using Optical Tweezers," PLOS ONE, Public Library of Science, vol. 5(8), pages 1-11, August.
    • Handle: RePEc:plo:pone00:0012224
    DOI: 10.1371/journal.pone.0012224
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    References listed on IDEAS

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    1. Matthew L. Walker & Stan A. Burgess & James R. Sellers & Fei Wang & John A. Hammer & John Trinick & Peter J. Knight, 2000. "Two-headed binding of a processive myosin to F-actin," Nature, Nature, vol. 405(6788), pages 804-807, June.
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