Abstract
Myosin V moves cargoes along actin filaments by walking hand over hand. Although numerous studies support the basic hand-over-hand model, little is known about the fleeting intermediate that occurs when the rear head detaches from the filament. Here we use submillisecond dark-field imaging of gold nanoparticle–labeled myosin V to directly observe the free head as it releases from the actin filament, diffuses forward and rebinds. We find that the unbound head rotates freely about the lever-arm junction, a trait that likely facilitates travel through crowded actin meshworks.
NOTE: In the version of this article initially published online, the length of the step shown in Figure 3e was mislabeled: it should be +25nm, not +24nm. In addition, the word "(right)" was erroneously included in the legend of Figure 2b. The errors have been corrected for all versions of the article.
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Change history
18 February 2007
NOTE: In the version of this article initially published online, the length of the step shown in Figure 3e was mislabeled: it should be +25nm, not +24nm. In addition, the word "(right)" was erroneously included in the legend of Figure 2b. The errors have been corrected for all versions of the article.
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Acknowledgements
We thank R. Vale (University of California, San Francisco) for the loan of the dark-field condenser and Z. Bryant and S. Churchman for insightful commentary. A.R.D. is a Jane Coffin Childs Postdoctoral Fellow. J.A.S. is supported by grant GM33289 from the US National Institutes of Health.
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Supplementary information
Supplementary Fig. 1
Expression of M5elc and M5cam (PDF 472 kb)
Supplementary Fig. 2
Sample trace showing alternating 53- and 21-nm steps (PDF 410 kb)
Supplementary Fig. 3
Step-size and dwell-time histograms (PDF 105 kb)
Supplementary Fig. 4
Sample axial and lateral displacements during the one-head-bound intermediate (PDF 140 kb)
Supplementary Fig. 5
Axial and lateral standard deviation histograms (PDF 455 kb)
Supplementary Fig. 6
Mathematical modeling of the one-head-bound state (PDF 94 kb)
Supplementary Fig. 7
Lateral displacement during the one-head-bound intermediate (PDF 144 kb)
Supplementary Fig. 8
M5elc free-head rebinding kinetics in the presence of 100 mM BDM (PDF 345 kb)
Supplementary Fig. 9
M5elc free-head rebinding kinetics in the presence of 50 mM free phosphate (PDF 113 kb)
Supplementary Video 1
M5elc labeled with a 40-nm gold particle walks on actin (MOV 546 kb)
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Dunn, A., Spudich, J. Dynamics of the unbound head during myosin V processive translocation. Nat Struct Mol Biol 14, 246–248 (2007). https://doi.org/10.1038/nsmb1206
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DOI: https://doi.org/10.1038/nsmb1206
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