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Journal of Muscle Research & Cell Motility

, Volume 12, Issue 4, pp 355–365 | Cite as

Discrimination of Ca2+-ATPase activity of the sarcoplasmic reticulum from actomyosin-type ATPase activity of myofibrils in skinned mammalian skeletal muscle fibres: distinct effects of cyclopiazonic acid on the two ATPase activities

  • Nagomi Kurebayashi
  • Yasuo Ogawa
Papers

Summary

We have developed a procedure to discriminate actomyosin-type ATPase activity from Ca2+-ATPase activity of sarcoplasmic reticulum (SR) in mechanically skinned fibres, determining simultaneously their Ca2+-induced tension and accompanying ATPase activity. When they were treated with an alkaline CyDTA-containing solution of low ionic strength which was reported to remove troponin C, the fibres showed a considerable amount of Ca2+-dependent ATPase activity, in spite of having little or no Ca2+-induced isometric tension. The residual ATPase activity is ascribed to the Ca2+-ATPase activity of SR, because it is completely abolished by 1% CHAPS treatment for 10 min. This conclusion is also supported by the finding that the Ca2+-dependence of the ATPase activity is very similar to that of Ca2+-ATPase of SR isolated from rabbit skeletal muscle, and that the estimated activity is consistent with the reported values of direct determinations. On the other hand, treatment with a detergent such as CHAPS or Triton X-100 removes SR activities (ATPase and Ca-uptake), leaving Ca2+-induced tension and actomyosin-type ATPase activity unchanged. This procedure indicated that the contribution of Ca2+-ATPase activity of SR may be minimal in total steady-state ATPase activity of mechanically skinned mammalian skeletal muscle fibres. Successive CyDTA and CHAPS treatments eliminated both Ca2+-induced tension and ATPase activity, which were recovered by the addition of troponin C. Using these procedures, we also examined the effect of cyclopiazonic acid (CPA) which was reported to be a specific inhibitor of Ca2+-ATPase of SR. Ca2+-ATPase activity of SR in skinned fibres was inhibited completely by 10 μm CPA and held to one-half by about 0.2 μm. This effect was only partially reversible. CPA at 10μm or higher concentrations showed Ca2+-sensitizing action on myofibrils, which was readily reversible. CPA at 3μm inhibited almost completely the Ca2+-ATPase activity of SR, while it had no effect on either actomyosin-type ATPase or isometric tension of myofibrils.

Keywords

Skeletal Muscle Ionic Strength ATPase Activity Sarcoplasmic Reticulum Specific Inhibitor 
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.

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References

  1. Duggan, P. F. &Martonosi, A. (1970) Sarcoplasmic reticulum IX. The permeability of sarcoplasmic reticulum membranes.J. Gen. Physiol. 56, 147–67.PubMedGoogle Scholar
  2. Ebashi, S. &Endo, M. (1986) Ca ion and muscle contraction.Prog. Biophys. Mol. Biol. 18, 123–83.Google Scholar
  3. Endo, M. (1977) Calcium release from the sarcoplasmic reticulum.Physiol. Rev. 57, 71–108.PubMedGoogle Scholar
  4. Goeger, D. R., Riley, R. T., Dorner, J. W. &Cole, R. J. (1988) Cyclopiazonic acid inhibition of the Ca2+-transport ATPase in rat skeletal muscle sarcoplasmic reticulum vesicles.Biochem. Pharmacol. 37, 978–81.PubMedGoogle Scholar
  5. Goldman, Y. E. (1987) Kinetics of the actomyosin ATPase in muscle fibers.Ann. Rev. Physiol. 49, 637–54.Google Scholar
  6. Guth, K. &Wojciechowski, R. (1986) Perfusion cuvette for the simultaneous measurement of mechanical, optical and energetic parameters of skinned muscle fibres.Pflugers Arch. 407, 552–7.PubMedGoogle Scholar
  7. Harafuji, H. &Ogawa, Y. (1980) Re-examination of the apparent binding constant of ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid with calcium around neutral pH.J. Biochem. 87, 1305–12.PubMedGoogle Scholar
  8. Homsher, E. (1987) Muscle enthalpy production and its relationship to actomyosin ATPase.Ann. Rev. Physiol. 49, 673–90.Google Scholar
  9. Inesi, G. (1985) Mechanism of calcium transport.Ann. Rev. Physiol. 47, 573–601.Google Scholar
  10. Kerrick, W. G. L., Secrist, D., Coby, R. &Lucas, S. (1976) Development of difference between red and white muscles in sensitivity to Ca2+ in the rabbit from embryo to adult.Nature 260, 440–1.PubMedGoogle Scholar
  11. Kurebayashi, N., Kodama, T. &Ogawa, Y. (1980) P1,P5-di(adenosine-5′)pentaphosphate (Ap5A) as an inhibitor of adenylate kinase in studies of fragmented sarcoplasmic reticulum from bullfrog skeletal muscle.J. Biochem. 88, 871–6.PubMedGoogle Scholar
  12. Kurebayashi, H., Ogawa, Y. &Harafuji, H. (1982) Effect of local anesthetics on calcium activated ATPase and its partial reaction with fragmented sarcoplasmic reticulum from bullfrog and rabbit skeletal muscle.J. Biochem. 92, 915–20.PubMedGoogle Scholar
  13. Kurebayashi, N. &Ogawa, Y. (1986) Characterization of increased Ca2+ efflux by quercetin from the sarcoplasmic reticulum in frog skinned skeletal muscle fibres.J. Muscle Res. Cell Motil. 7, 142–50.PubMedGoogle Scholar
  14. Kurebayashi, N. &Ogawa, Y. (1988) Increase by trifluoperazine in calcium sensitivity of myofibrils in a skinned fibre from frog skeletal muscle.J. Physiol. 403, 407–24.PubMedGoogle Scholar
  15. Martonosi, A. N. &Beeler, T. J. (1983) Mechanism of Ca2+ transport by sarcoplasmic reticulum. InHandbook of Physiology, Section 10, Skeletal Muscle (edited by Peachey, L. D., Adrian, R. H. & Geiger, S. R.) pp. 417–85. Bethesda: American Physiological Society.Google Scholar
  16. Morimoto, S. &Ohtsuki, I. (1987) Ca2+- and Sr2+-sensitivity of the ATPase activity of rabbit skeletal myofibrils: effect of complete substitution of troponin C with cardiac troponin C, calmodulin, and parvalbumins.J. Biochem. 101, 291–301.PubMedGoogle Scholar
  17. Ogawa, Y. &Kurebayashi, N. (1982) ATP-ADP exchange reaction by fragmented sarcoplasmic reticulum from bullfrog skeletal muscle.J. Muscle Res. Cell Motil. 3, 39–56.PubMedGoogle Scholar
  18. Ogawa, Y. (1985) Sarcoplasmic reticulum: Ca-pump. InHandbook of Physiological Sciences, Vol. 4,Physiology of Skeletal, Cardiac and Smooth Muscles (edited by Tomita, T. & Sugi, H.) pp. 91–101. Tokyo: Igaku Shoin (in Japanese).Google Scholar
  19. Ogawa, Y. (1988) Comparative aspects of the mechanisms of energy transduction in sarcoplasmic reticulum between rabbit and frog skeletal muscle. InThe Roots of Modern Biochemistry (edited by Kleinkauf, H. Von Döhren, H. & Jaenicke, L.) Pp. 747–58. Berlin, New York: Walter de Gruyter.Google Scholar
  20. Ogawa, Y. &Kurebayashi, N. (1989) Modulations by drugs of the relationship between calcium binding to troponin C and tension. InMuscle Energetics (edited by Paul, R., Elzinga, G. & Yamada, K.) pp. 75–86. New York: Alan R. Liss.Google Scholar
  21. Seidler, N. W., Jona, I., Vegh, M. &Martonosi, A. (1989) Cyclopiazonic acid is a specific inhibitor of the Ca2+-ATPase of sarcoplasmic reticulum.J. Biol. Chem. 264, 17816–23.PubMedGoogle Scholar
  22. Stephenson, D. G. &Williams, D. A. (1981) Calcium-activated force responses in fast- and slow-twitch skinned muscle fibres of rat at different temperatures.J. Physiol. 317, 281–302.PubMedGoogle Scholar
  23. Stephenson, D. G., Stewart, A. W. &Wilson, G. J. (1989) Dissociation of force from myofibrillar MgATPase and stiffness at short sarcomere lengths in rat and toad skeletal muscle.J. Physiol. 410, 351–66.PubMedGoogle Scholar
  24. Takagi, A. &Endo, M. (1977) Guinea pig soleus and extensor digitorum longus: a study on single-skinned fibers.Exp. Neurol. 55, 95–101.PubMedGoogle Scholar
  25. Yates, L. D. &Greaser, M. L. (1983) Quantitative determination of myosin and actin in rabbit skeletal muscle.J. Mol. Biol. 168, 123–41.PubMedGoogle Scholar

Copyright information

© Chapman and Hall Ltd 1991

Authors and Affiliations

  • Nagomi Kurebayashi
    • 1
  • Yasuo Ogawa
    • 1
  1. 1.Department of PharmacologyJuntendo University School of MedicineTokyoJapan

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