Advertisement

Electrostatic Potential Around Actin

  • Toshio Ando
  • Naohiro Kobayashi
  • Eisuke Munekata
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 332)

Abstract

We presume that tension of contracting muscle originates from electrostatic force experienced by actin and myosin. We suppose that a high-energy state of myosin-ADP-Pi interacts with actin, transferring the stored energy to actin, and that the actin excited in this way develops around itself electric field which exerts sliding force against charged myosin heads. To explore the idea, first we conjectured how electric charges on actin produce electric field in the axial direction, and second we experimentally examined electrostatic circumstances around actin in Solution and in muscle fibers by optimizing diffusion-enhanced fluorescence energy transfer. In the experiments, Tb ion, which has a long excited-state lifetime, was used as donor. To introduce Tb to actin, Tb-DTPA-phalloin and Tb-DTPA-maleimide were synthesized. As acceptors with electric charges (Za = -3 to +2), rhodamine B that was conjugated with various amino acids or their derivatives was used. The fluorescence energy transfer efficiency (ET) was estimated from the shortening in the lifetime of Tb. The electrostatic circumstances around actin were inferred from the ET-Za relation. When Tb was introduced at Cys-374 of actin, the Tb-site was found in negative electric potential. S-l binding to the labeled actin neutralized the electric potential almost completely. Tb-DTPA-phalloin bound to actin seemed to reside in the vicinity of tryptophan residue(s). Electric potential around the phalloin site was negative. S-l binding to the actin slightly reduced the negativity. In glycerinated fibers in the rigor state, the phalloin site was again found in negative potential. When fibers were transferred from an ADP-rigor solution to an active solution, the negative electric potential was neutralized to some extent. The direction of this change could not be explained by detachment of crossbridges from actin, since the detachment should have given an opposite direction of changes in the electric potential. Thus, this observation may indicate that electric potential characteristic of the active state occurs at actin surfaces.

Keywords

Electric Charge Electric Potential Actin Filament Myosin Head Energy Transfer Efficiency 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Vale, R.D., Schnapp, B.J., Mitchinson, T., Steuer, E., Reese, T.S. & Sheetz, M.P. Cell 43, 623–632 (1985).PubMedCrossRefGoogle Scholar
  2. 2.
    Endow, S.A., Henikoff, S. & Soler-Niedziela, L. Nature 345, 81–83 (1990).PubMedCrossRefGoogle Scholar
  3. 3.
    Walker, R.A., Salmon, E.D. & Endow, S.A. Nature 347, 780–782 (1990).PubMedCrossRefGoogle Scholar
  4. 4.
    McDonald, H.B., Stewart, R.J. & Goldstein, L.S.B. Cell 63, 1159–1165 (1990).PubMedCrossRefGoogle Scholar
  5. 5.
    Schliwa, M., Shimizu, T., Vale, R.D. & Euteneuer, U. J. Cell Biol. 112, 1199–1203 (1991).PubMedCrossRefGoogle Scholar
  6. 6.
    Malik, F. & Vale, R.D. Nature 347, 713–714 (1990).PubMedCrossRefGoogle Scholar
  7. 7.
    Mornet, D., Bertrand, R., Pantel, P., Audemard, E. & Kassab, R. Nature 292, 301–306 (1981).PubMedCrossRefGoogle Scholar
  8. 8.
    Sutoh, K. Biochemistry 22, 1579–1585 (1983).PubMedCrossRefGoogle Scholar
  9. 9.
    Yamamoto, K. Biochemistry 28, 5573–5577 (1989).PubMedCrossRefGoogle Scholar
  10. 10.
    Katoh, T. & Morita, F. J. Biochem. 96, 1223–1230 (1984).PubMedGoogle Scholar
  11. 11.
    Highsmith, S. Biochemistry 29, 10690–10694 (1990).PubMedCrossRefGoogle Scholar
  12. 12.
    Elliot, G.F., Rome, E. & Spencer, M. Nature 226, 417–420 (1970).CrossRefGoogle Scholar
  13. 13.
    Iwazumi, T. in Cross-Bridge Mechanism in Muscle Contraction (eds. Sugi & Pollack, Univ. of Tokyo Press, Tokyo, 1979).Google Scholar
  14. 14.
    Wieland, T., Hollosi, M., & Nassal, M. Liebigs Ann. Chem. 1983, 1533–1540 (1983).CrossRefGoogle Scholar
  15. 15.
    Oosawa, F., Asakura, S. & Ooi, T. Suppl Prog. Theor. Phys. 17, 14–34 (1961).CrossRefGoogle Scholar
  16. 16.
    Yanagida, T. J. Muscle Res. Cell Motility 6, 43–52 (1985).CrossRefGoogle Scholar
  17. 17.
    Erickson, H.P. J. Mol. Biol. 206, 465–474 (1989).PubMedCrossRefGoogle Scholar
  18. 18.
    Fujime, S., Ishiwata, S. & Maeda, T. Biochim. Biophys. Acta 283, 351–363 (1972).PubMedCrossRefGoogle Scholar
  19. 19.
    Mihashi, K., Yoshimura, H., Nishio, T., Ikegami, A. & Kinoshita, K. Jr. J. Biochem. 93, 1705–1707 (1983).PubMedGoogle Scholar
  20. 20.
    Stokes, D.L. & DeRosier, DJ. J. Cell Biol. 104, 1005–1017 (1987).PubMedCrossRefGoogle Scholar
  21. 21.
    Kabsch, W., Mannherz, H.G., Suck, D., Pai, E.F. & Holmes, K.C. Nature 347, 37–44 (1990).PubMedCrossRefGoogle Scholar
  22. 22.
    Wensel, T.G. & Meares, C.F. Biochemistry 22, 6247–6254 (1983).CrossRefGoogle Scholar
  23. 23.
    Vandekerckhove, J., Deboben, A., Nassal, M. & Wieland, T. EMBO J. 4, 2815–2818 (1985).PubMedGoogle Scholar
  24. 24.
    Holmes, K.C, Popp, D., Gebhard, W. & Kabsch, W. Nature, 347, 44–49 (1990).PubMedCrossRefGoogle Scholar
  25. 25.
    Sutoh, K., Ando, M., Sutoh, K. & Yano-Toyoshima, Y. Proc. Natl. Acad. Sci. USA 88, 7711–7714 (1991).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • Toshio Ando
    • 1
  • Naohiro Kobayashi
    • 2
  • Eisuke Munekata
    • 2
  1. 1.Department of Physics Faculty of ScienceKanazawa UniversityJapan
  2. 2.Department of Applied BiochemistryTsukuba UniversityTsukubaJapan

Personalised recommendations