Skip to main content
SpringerLink
Log in

Factors Affecting Filament Velocity in In Vitro Motility Assays and their Relation to Unloaded Shortening Velocity in Muscle Fibers

  • Chapter
Mechanism of Myofilament Sliding in Muscle Contraction

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 332))

Abstract

The measurement of fluorescently labeled actin filament movement driven by mechanoenzymes (e.g. myosin) is an important methodology for the study of molecular motors. It is assumed that the filament velocity, Vf, is analogous to the unloaded shortening velocity, Vu, seen in muscle fibers. To evaluate this assumption we compared Vf to literature values for Vu with regard to the effects of [ATP], [ADP], [Pi], pH, ionic strength (10–150 mM) and temperature (15–30°C). Vf and Vu are quantitatively similar with respect to the effects of substrate and product concentrations and temperatures > 20°C. However, Vf is more sensitive to decreases in pH and temperatures < 20°C than is Vu. At ionic strengths of 50–150 mM, Vf and Vu exhibit similar ionic strength dependencies (decreasing with ionic strength). At ionic strengths < 50 mM, Vf is markedly reduced. Thus while Vf is a good analogue for Vu under certain conditions (elevated ionic strength and temperatures > 20°C), under others it is not. The results of motility assays must be cautiously interpreted.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 39.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Homsher, E. & Millar, N. Ann. Rev. Physiol. 52, 875–896 (1990).

    Article  CAS  Google Scholar 

  2. Brenner, B., M. Schoenberg, M., Chalovich, J.M., Greene, L.E. & Eisenberg, E. Proc. Natl. Acad. Sci. USA 79, 7288–7291 (1982).

    Article  PubMed  CAS  Google Scholar 

  3. Hibberd, M., Dantzig, J.A., Trentham, D.R. & Goldman, Y.E. Science 228, 1317–1319 (1985).

    Article  PubMed  CAS  Google Scholar 

  4. Pate, E. & Cooke, R. J. Muscle Res. Cell Motility 10, 181–196 (1989).

    Article  CAS  Google Scholar 

  5. Siemankowski, R. F, Wiseman, M.O. & White, H.D. Proc. Natl. Acad. Sci. USA 82, 658–662 (1985).

    Article  PubMed  CAS  Google Scholar 

  6. Cooke, R. & Pate, E. Biophys. J. 48, 789–798 (1985).

    Article  PubMed  CAS  Google Scholar 

  7. Harada, Y., Noguchi, A., Kishino, A. & Yanagida, T. Nature 326, 805–808 (1987).

    Article  PubMed  CAS  Google Scholar 

  8. Kron, SJ. & Spudich, J.A. Proc. Natl. Acad. Sci. USA 83, 6272–6276 (1986).

    Article  PubMed  CAS  Google Scholar 

  9. Sheetz, M. P. & Spudich, J.A. Nature 303, 31–35 (1983).

    Article  PubMed  CAS  Google Scholar 

  10. Homsher, E., Wang, F. & Sellers, J. Am. J. Physiol. (Cell) in press

    Google Scholar 

  11. Fabiato, A. & Fabiato, F. J. Physiol. (Paris) 75, 463–505 (1979).

    CAS  Google Scholar 

  12. Ferenczi, M.A., Goldman, Y.E. & Simmons, R.M. J. Physiol. (Lond.) 350, 519–543 (1984).

    CAS  Google Scholar 

  13. Sheetz, N.P., Chasan, R. & Spudich, J.A. J. Cell Biol. 99, 1867–1871 (1984).

    Article  PubMed  CAS  Google Scholar 

  14. Umemoto, S. & Sellers, J.R. J. Biol. Chem. 265, 14864–14869 (1990).

    PubMed  CAS  Google Scholar 

  15. Warshaw, D.M., Desrosiers, J.M., Work, S.S. & Trybus, K.M. J. Cell Biol. 111, 453–463 (1990).

    Article  PubMed  CAS  Google Scholar 

  16. Eisenberg, E. & Keilley, W. J. Biol. Chem. 249, 4742–4748 (1974)

    PubMed  CAS  Google Scholar 

  17. Pate, E., Nakamaya, K., Franks-Skiba, K., Yount, R.G. & Cooke, R. Biophys. J. 59, 598–605 (1991).

    Article  PubMed  CAS  Google Scholar 

  18. Cooke, R., Franks, K., Luciani, G. & Pate, E. J. Physiol. (Lond.) 395, 77–97 (1988).

    CAS  Google Scholar 

  19. Gulati, J. & Podolsky, R.J. J. Gen. Physiol. 78, 233–257 (1981).

    Article  PubMed  CAS  Google Scholar 

  20. Takiguchi, K., Hayashi, H., Kurimoto, E. & Higashi-Fujime, S. J. Biochem. 107, 671–679 (1990).

    PubMed  CAS  Google Scholar 

  21. Vale, R.D. & Oosawa, F. Adv. Biophys. 26, 97–134 (1990).

    Article  PubMed  CAS  Google Scholar 

  22. Uyeda, T.Q.P., Krön, SJ. & Spudich, J.A. J. Mol. Biol. 214, 699–710 (1990).

    Article  PubMed  CAS  Google Scholar 

  23. Tesi, C., Kitagishi, K., Travers, F. & Barman, T. Biochemistry 30, 4061–4067 (1991).

    Article  PubMed  CAS  Google Scholar 

  24. Ranatunga, K.W. J. Physiol. (Lond.) 329, 465–483 (1982).

    CAS  Google Scholar 

  25. Stein, R.B., Gordon, T. & Shriver, J. Biophys. J. 40, 97–107 (1982).

    Article  PubMed  CAS  Google Scholar 

  26. Sweeney, H.L., Kushmerick, M.J., Mabuchi, K., Sreter, F. & Gergely, J. J. Biol. Chem. 263, 9034–9039 (1988).

    PubMed  CAS  Google Scholar 

  27. Toyoshima, Y.Y., Kron, SJ. & Spudich, J.A. Proc. Natl Acad. Sci. USA 87, 7130–7134 (1990).

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Section of Cellular and Molecular Motility, Laboratory of Molecular Cardiology, Building 10, Rm. 8N202, NIH, Bethesda, Maryland, 20892, USA

    Fei Wang & James Sellers

  2. Department of Physiology, School of Medicine, UCLA, Los Angeles, California, 90024, USA

    Earl Homsher

Authors
  1. Earl Homsher
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. James Sellers
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Teikyo University, Tokyo, Japan

    Haruo Sugi

  2. University of Washington, Seattle, Washington, USA

    Gerald H. Pollack

Rights and permissions

Reprints and permissions

Copyright information

© 1993 Springer Science+Business Media New York

About this chapter

Cite this chapter

Homsher, E., Wang, F., Sellers, J. (1993). Factors Affecting Filament Velocity in In Vitro Motility Assays and their Relation to Unloaded Shortening Velocity in Muscle Fibers. In: Sugi, H., Pollack, G.H. (eds) Mechanism of Myofilament Sliding in Muscle Contraction. Advances in Experimental Medicine and Biology, vol 332. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2872-2_27

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-2872-2_27

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-6245-6

  • Online ISBN: 978-1-4615-2872-2

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation