Force measurements by micromanipulation of a single actin filament by glass needles (original) (raw)

Nature volume 334, pages 74–76 (1988)Cite this article

Abstract

Single actin filaments (∼7nm in diameter) labelled with fluorescent phalloidin can be clearly seen by video-fluorescence microscopy1. This technique has been used to observe motions of single filaments in solution and in several in vitro movement assays1–5. In a further development of the technique, we report here a method to catch and manipulate a single actin filament (F-actin) by glass microneedles under conditions in which external force on the filament can be applied and measured. Using this method, we directly measured the tensile strength of a filament (the force necessary to break the bond between two actin monomers) and the force required for a filament to be moved by myosin or its proteolytic fragment bound to a glass surface in the presence of ATP. The first result shows that the tensile strength of the F-actin–phalloidin complex is comparable with the average force exerted on a single thin filament in muscle fibres during isometric contraction. This force is increased only slightly by tropomyosin. The second measurement shows that the myosin head (subfragment-1) can produce the same ATP-dependent force as intact myosin. The magnitude of this force is comparable with that produced by each head of myosin in muscle during isometric contraction.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Yanagida, T., Nakase, M., Nishiyama, K. & Oosawa, F. Nature 307, 58–60 (1984).
    Article ADS CAS Google Scholar
  2. Honda, H., Nagashima, H. & Asakura, S. J. molec. Biol. 191, 113–133 (1986).
    Article Google Scholar
  3. Kron, S. J. & Spudich, J. A. Proc. natn. Acad. Sci. U.S.A. 83, 62–72 (1986).
    Article Google Scholar
  4. Harada, Y., Noguchi, A., Kishino, A. & Yanagida, T. Nature 326, 805–808 (1987).
    Article ADS CAS Google Scholar
  5. Yano-Toyoshima, Y. et al. Nature 328, 536–539 (1986).
    Article Google Scholar
  6. Dancker, P., Low, I., Hasselbach, W. & Wieland, Th. Biochim. biophys. Acta 400, 407–414 (1975).
    Article CAS Google Scholar
  7. Kamimura, S. & Takahashi, K. Nature 293, 266–268 (1981).
    Article Google Scholar
  8. Oosawa, F. Biorheology 14, 11–19 (1977).
    Article CAS Google Scholar
  9. McLachlan, A. D. & Stewart, M., J. molec. Biol. 98, 293–304 (1975).
    Article CAS Google Scholar
  10. Phillips, G. N. Jr, Fillers, J. P. & Cohen, C. J. molec. Biol. 192, 111–131 (1986).
    Article CAS Google Scholar
  11. Prochniewicz-Nakayama, E., Yanagida, T. & Oosawa, F. J. Cell Biol. 97, 1663–1667 (1983).
    Article CAS Google Scholar
  12. Yano, M., Yamamoto, Y. & Shimizu, H. Nature 299, 557–559 (1982).
    Article ADS CAS Google Scholar
  13. Sheetz, M. P. & Spudich, J. A. Nature 303, 31–35 (1983).
    Article ADS CAS Google Scholar
  14. Hynes, T., Block, S. M., White, B. T. & Spudich, J. A. Cell 48, 953–963 (1987).
    Article CAS Google Scholar
  15. Lanzetta, P. A., Alvarez, L. J., Reinach, P. S. & Candia, O. A. Analyt. Biochem. 100, 95–97 (1979).
    Article CAS Google Scholar
  16. Kamimura, S. Appl. Optics 26, 3425–3427 (1987).
    Article ADS CAS Google Scholar
  17. Gelles, J., Schnapp, B. J. & Sheetz, M. P. Nature 331, 450–453 (1988).
    Article ADS CAS Google Scholar
  18. Yoneda, M. J. exp. Biol. 37, 461–468 (1960).
    Google Scholar
  19. Margossian, S. S. & Lowey, S. J. molec. Biol. 74, 313–330 (1973).
    Article CAS Google Scholar

Download references

Author information

Authors and Affiliations

  1. Department of Biophysical Engineering, Faculty of Engineering Science, Osaka University, Toyonaka, Osaka, Japan
    Akiyoshi Kishino & Toshio Yanagida

Authors

  1. Akiyoshi Kishino
    You can also search for this author inPubMed Google Scholar
  2. Toshio Yanagida
    You can also search for this author inPubMed Google Scholar

Rights and permissions

About this article

Cite this article

Kishino, A., Yanagida, T. Force measurements by micromanipulation of a single actin filament by glass needles.Nature 334, 74–76 (1988). https://doi.org/10.1038/334074a0

Download citation