Advertisement

Photochemical Cleavage of Myosin Heavy Chain and the Effect on the Interaction with Actin

  • Yoh Okamoto
  • Christine Cremo
Chapter
  • 101 Downloads
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 332)

Abstract

Myosin from rabbit skeletal muscle has been photochemically cleaved in the presence of vanadate ion and ATP. Two cleavage sites termed V1 and V2 within the S1 head region were studied. In the presence of magnesium ion both sites were cleaved, but in the absence of divalent cation cleavage only occurred at the V2 site. V2 cleaved myosin had higher K+-EDTA-ATPase and actin activated Mg2+-ATPase activity than V1, V2 cleaved myosin. Immunochemical characterization shows that the photochemical cleavage is more specific than that of proteolytic cleavage since breakdown of light chains was not observed for the photochemical method. This must be one of the best ways to prepare a single site cleaved myosin for the study of molecular mechanism of sliding.

Keywords

ATPase Activity Heavy Chain Myosin Heavy Chain Head Region Myosin Head 
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.
    Korn, E.D. & Hammer, J.A.III Ann. Rev. Biophys. Biophys. Chem. 17, 23–45 (1988).CrossRefGoogle Scholar
  2. 2.
    Pollard, T.D. et al. Ann. Rev. Physiol. 53, 653–681 (1991).CrossRefGoogle Scholar
  3. 3.
    Kabsh, W. et al. Nature 347, 37–44 (1990).CrossRefGoogle Scholar
  4. 4.
    Okamoto, Y. & Yount, R.G. Proc. Natl. Acad. Sci. USA 82, 1575–1579 (1985).PubMedCrossRefGoogle Scholar
  5. 5.
    Mahmood, R. & Yount, R.G. J. Biol. Chem. 259, 12956–12959 (1984).PubMedGoogle Scholar
  6. 6.
    Okamoto, Y. & Sekine, T. J. Biol. Chem. 262, 7851–7854 (1987).Google Scholar
  7. 7.
    Kielley, W.W. & Bradley, L.B. J. Biol. Chem. 218, 653–659 (1956).PubMedGoogle Scholar
  8. 8.
    Spudich, J.A. & Watt, S. J. Biol. Chem. 246, 4866–4871 (1971).PubMedGoogle Scholar
  9. 9.
    Goodno, C.C. Methods Enzymol. 85, 116–123 (1982).PubMedCrossRefGoogle Scholar
  10. 10.
    Fiske, C.H. & SubbaRow, Y. J. Biol. Chem. 66, 375–400 (1925).Google Scholar
  11. 11.
    Laemmli, U.K. Nature 227, 680–685 (1970).PubMedCrossRefGoogle Scholar
  12. 12.
    Okamoto, Y. et al. Nature 324, 78–80 (1986).PubMedCrossRefGoogle Scholar
  13. 13.
    Lee-Eiford, A. et al. J. Biol. Chem. 261, 2337–2342 (1986).PubMedGoogle Scholar
  14. 14.
    Grammer, J.C. et al. Biochemistry 27, 8408–8415 (1988).PubMedCrossRefGoogle Scholar
  15. 15.
    Cremo, C.R. et al. Biochemistry 27, 8415–8420 (1988).PubMedCrossRefGoogle Scholar
  16. 16.
    Mocz, G. Eur. J. Biochem. 179, 373–378 (1989).PubMedCrossRefGoogle Scholar
  17. 17.
    Cremo, C.R. et al. Biochemistry 29, 7982–7990 (1990).PubMedCrossRefGoogle Scholar
  18. 18.
    Mühlrad, A. et al. Biochemistry 30, 958–965 (1991).PubMedCrossRefGoogle Scholar
  19. 19.
    Wells, J.A. & Yount, R.G. Methods Enzymol. 85, 93–116 (1982).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • Yoh Okamoto
    • 1
  • Christine Cremo
    • 2
  1. 1.Division of Bioengineering, Department of Applied ChemistryMuroran Institute of TechnologyMuroranJapan
  2. 2.Biochemistry and Biophysics DepartmentWashington State UniversityPullmanUSA

Personalised recommendations