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Substeps within the 8-nm step of the ATPase cycle of single kinesin molecules

Nature Cell Biology volume 3, pages 425–428 (2001)Cite this article

Abstract

Kinesin is a molecular motor that moves processively1,2,3,4 by regular 8-nm steps along microtubules5,6,7,8,9,10,11. The processivity of this movement is explained by a hand-over-hand model in which the two heads of kinesin work in a coordinated manner. One head remains bound to the microtubule while the other steps from the αβ-tubulin dimer behind the attached head to the dimer in front. The overall movement is 8 nm per ATPase cycle9,10,11,12,13. To investigate elementary processes within the 8-nm step, we have developed a new assay that resolves nanometre displacements of single kinesin molecules with microsecond accuracy. Our data show that the 8-nm step can be resolved into fast and slow substeps, each corresponding to a displacement of ∼4 nm. The substeps are most probably generated by structural changes in one head of kinesin, leading to rectified forward thermal motions of the partner head14. It is also possible that the kinesin steps along the 4-nm repeat of tubulin monomers.

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Figure 1: Schematic diagram of laser trapping nanometry system (not to scale).
Figure 2: Kinesin-driven bead displacement and response time.
Figure 3: Rising phases of single 8 nm steps.
Figure 4: Analysis of the rising phase of the 8 nm step.

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References

  1. Vale, R. D., Reese, T. S. & Sheetz, M. P. Cell 42, 39–50 (1985).

    Article  CAS  Google Scholar 

  2. Howard, J., Hudspeth, A. J. & Vale, R. D. Nature 342, 154–158 (1989).

    Article  CAS  Google Scholar 

  3. Gilbert, S. P., Webb, M. R., Brune, M. & Johnson, K. A. Nature 373, 671–676 (1995).

    Article  CAS  Google Scholar 

  4. Hackney, D. D. Nature 377, 448–450 (1995).

    Article  CAS  Google Scholar 

  5. Higuchi, H., Muto, E., Inoue, Y. & Yanagida, T. Proc. Natl Acad. Sci. USA 94, 4395–4400 (1997).

    Article  CAS  Google Scholar 

  6. Crevel, I., Carter, N., Schliwa, M. & Cross, R. EMBO J. 18, 5863–5872 (1999).

    Article  CAS  Google Scholar 

  7. Coppin, C. M., Finer, J. T., Spudich, J. A. & Vale, R. D. Proc. Natl Acad. Sci. USA 93, 1913–1917 (1996).

    Article  CAS  Google Scholar 

  8. Svoboda, K., Schmidt, C. F., Schnapp, B. J. & Block, S. M. Nature 365, 721–727 (1993).

    Article  CAS  Google Scholar 

  9. Kojima, H., Muto, E., Higuchi, H. & Yanagida, T. Biophys. J. 73, 2012–2022 (1997).

    Article  CAS  Google Scholar 

  10. Schnitzer, M. J. & Block, S. M. Nature 388, 386–390 (1997).

    Article  CAS  Google Scholar 

  11. Visscher, K., Schnitzer, M. J. & Block, S. M. Nature 400, 184–189 (1999).

    Article  CAS  Google Scholar 

  12. Hua, W., Young, E. C., Fleming, M. L. & Gelles, J. Nature 388, 390–393 (1997).

    Article  CAS  Google Scholar 

  13. Coy, D. L., Wagenbach, M. & Howard, J. J. Biol. Chem. 274, 3667–3671 (1999).

    Article  CAS  Google Scholar 

  14. Rice, S. et al. Nature 402, 778–784 (1999).

    Article  CAS  Google Scholar 

  15. Allersma, M. W., Gittes, F., deCastro, M. J., Stewart, R. J. & Schmidt, C. F. Biophys. J. 74, 1074–1085 (1998).

    Article  CAS  Google Scholar 

  16. Svoboda, K. & Block, S. M. Cell 77, 773–784 (1994).

    Article  CAS  Google Scholar 

  17. Meyhofer, E. & Howard, J. Proc. Natl Acad. Sci. USA 92, 574–578 (1995).

    Article  CAS  Google Scholar 

  18. Kudo, S., Magariyama, Y. & Aizawa, S. Nature 346, 677–680 (1990).

    Article  CAS  Google Scholar 

  19. Hirose, K., Lockhart, A., Cross, R. A. & Amos, L. A. Proc. Natl Acad. Sci. USA 93, 9539–9544 (1996).

    Article  CAS  Google Scholar 

  20. Walker, R. A. Proc. Natl Acad. Sci. USA 92, 5960–5964 (1995).

    Article  CAS  Google Scholar 

  21. Larcher, J. C., Boucher, D., Lazereg, S., Gros, F. & Denoulet, P. J. Biol. Chem. 271, 22117–22124 (1996).

    Article  CAS  Google Scholar 

  22. Tucker, C. & Goldstein, L. S. B. J. Biol. Chem. 272, 9481–9488 (1997).

    Article  CAS  Google Scholar 

  23. Nogales, E., Wolf, S. G. & Downing, K. H. Nature 391, 199–203 (1998).

    Article  CAS  Google Scholar 

  24. Nogales, E., Whittaker, M., Milligan, R. A. & Downing, K. H. Cell 96, 79–88 (1999).

    Article  CAS  Google Scholar 

  25. Nishiyama, M., Higuchi, H., Muto, E., Inoue, Y. & Yanagida, T. Biophys. J. 78, 122A (2000).

    Google Scholar 

Download references

Acknowledgements

We thank K. Kitamura and A. Ishijima for technical suggestions, Y. Ishii and other colleagues of Single Molecule Processes Project, JST and Osaka University for valuable discussions, and H. Kojima, Y. E. Goldman, F. Brozovich, Y. Ishii Jr, J. West and S. A. Endow for critically reading the manuscript. This work was partially supported by JSPS Research Fellowships for Young Scientists (M.N.), the Japan Ministry of Education, Science, Sport and Culture (H.H.), and by Yanagida BioMotron Project, ERATO, JST. A preliminary report of this investigation has been presented previously25.

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Authors and Affiliations

  1. Department of Biophysical Engineering, Osaka University 1-3, Machikaneyama, Toyonaka, 560-8531, Osaka, Japan

    Masayoshi Nishiyama & Toshio Yanagida

  2. Form and Function, PRESTO, JST, 2-4-14, Senba-Higashi, Mino, 562-0035, Osaka, Japan

    Etsuko Muto

  3. Single Molecule Processes Project, ICORP, JST, 2-4-14, Senba-Higashi, Mino, 562-0035, Osaka, Japan

    Yuichi Inoue & Toshio Yanagida

  4. Department of Physiology and Biosignaling, Graduate School of Medicine, Osaka University 2-2, Yamadaoka, Suita, 565-0871, Osaka, Japan

    Toshio Yanagida

  5. Department of Metallurgy, Graduate School of Engineering, Tohoku University, Sendai, 980-8579, Japan

    Hideo Higuchi

  6. Center of Interdisciplinary Research, Tohoku University, Sendai, 980-8579, Japan

    Hideo Higuchi

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  1. Masayoshi Nishiyama
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  2. Etsuko Muto
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  5. Hideo Higuchi
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Correspondence to Toshio Yanagida.

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Nishiyama, M., Muto, E., Inoue, Y. et al. Substeps within the 8-nm step of the ATPase cycle of single kinesin molecules. Nat Cell Biol 3, 425–428 (2001). https://doi.org/10.1038/35070116

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