Journal of Muscle Research & Cell Motility

, Volume 4, Issue 5, pp 543–556 | Cite as

Generation of tension by glycerol-extracted vertebrate skeletal muscle fibres in the absence of calcium

  • H. D. Loxdale
  • R. T. Tregear


When a small bundle of glycerol-extracted fibres from either frog, tortoise or rabbit skeletal muscle was first exposed to high MgATP (5mm) in the absence of Ca2+ (<1nm) and at low ionic strength (<0.11) at 20° C, it produced a single sharp transient contraction followed by a lower maintained isometric tension. The maintained tension was investigated further in rabbit psoas fibres. Ca2+-free tension was dependent on the ionic strength in the range 0.04–0.10, on the temperature in the range 6–20° C and the free Mg2+ in the range 0–6mm. It was promoted by low ionic strength, low Mg2+ and high temperature, and was unaffected by varying the MgATP2− in the range 0.4–4mm and by adding ATP regenerating components. A separate regime of tension generation was detected at MgATP2− concentrations of less than 0.1mm, in which MgATP2− concentration was critical. The results are interpreted on the assumption that binding of Mg2+ to some component of the regulatory system is necessary to maintain its inhibitory effect in the absence of Ca2+. Ionic strength and temperature, on the other hand, may affect actomyosin directly.


Calcium Skeletal Muscle Muscle Fibre Ionic Strength Regulatory System 
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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  1. ABBOTT, R. H. (1966) The relaxation of glycerinated rabbit psoas muscle fibres by EGTA.J. Physiol. 186, 115–6.Google Scholar
  2. ABBOTT, R. J. & LEECH, A. R. (1973) Persistence of adenylate kinase and other enzymes in glycerol-extracted muscle.Pflügers Arch. 344, 233–43.Google Scholar
  3. BAGSHAW, C. R. (1977) On the location of the divalent metal binding sites and the light chain subunits of vertebrate myosin.Biochemistry 16, 59–67.Google Scholar
  4. CHIN, T. K. & ROWE, A. J. (1982) Biochemical properties of native myosin filaments.J. Musc. Res. Cell. Motility 3, 118.Google Scholar
  5. DONALDSON, S. K. B. & KERRICK, W. G. L. (1975) Characterisation of the effects of Mg2+ on Ca2+ and Sr2+-activated tension generation of skinned skeletal muscle fibres.J. gen. Physiol. 66, 417–44.Google Scholar
  6. EATON, B. L., KOMINZ, D. R. & EISENBERG, E. (1975) Correlation between the inhibition of the acto-heavy meromyosin ATPase and the binding of tropomyosin to F-actin: Effects of Mg2+, KCl, troponin-I, and troponin-C.Biochemistry 14, 2718–25.Google Scholar
  7. EMBRY, R. & BRIGGS, A. H. (1966) Factors affecting contraction and relaxation in dog glycerinated cardiac fibres.Am. J. Physiol. 210, 826–30.Google Scholar
  8. FELDHAUS, P., FROHLICH, T., GOODY, R. S., ISAKOV, M. & SCHIRMER, R. H. (1975) Synthetic inhibitors of adenylated kinases in the assays for ATPases and phosphokinases.Eur. J. Biochem. 57, 197–204.Google Scholar
  9. FERENCZI, M. A., SIMMONS, R. M. & SLEEP, J. A. (1982) General considerations of crossbridge models in relation to the dependence on MgATP concentration of mechanical parameters of skinned fibres from frog muscle. InBasic Biology of Muscles: a Comparative Approach (edited by TWAROG, B. M., LEVINE, R. J. C. and DEWEY, M. M.), pp. 91–107. New York: Raven Press.Google Scholar
  10. FUCHS, F. & BLACK, B. (1980) The effects of magnesium on the binding of calcium ions to glycerinated rabbit psoas muscle fibres.Biochim. biophys. Acta 622, 52–62.Google Scholar
  11. GORDON, A. M., GODT, R. E., DONALDSON, S. K. B. & HARRIS, C. E. (1973) Tension of skinned frog muscle fibres in solutions of varying ionic strength and neutral salt composition.J. gen. Physiol. 62, 550–74.Google Scholar
  12. GREENE, L. E. & EISENBERG, E. (1980) Co-operative binding of myosin subfragment-1 to the actin-troponin-tropomyosin complex.Proc. natn. Acad. Sci. 77, 2616–20.Google Scholar
  13. GRIFFITHS, P. J., KUHN, H. J. & RÜEGG, J. C. (1979) Activation of the contractile system of insect fibrillar muscle at very low concentrations of Mg2+ and Ca2+.Pflügers Arch. 382, 155–63.Google Scholar
  14. GUNTHER, T. (1967) Intracellular magnesium ion concentration.Z. Naturforsch. 226, 149–54.Google Scholar
  15. HIGHSMITH, S. (1977) The effects of temperature and salts on myosin subfragment-1 and F-actin complexes.Archs Biochem. Biophys. 180, 404–8.Google Scholar
  16. HOMSHER, E., BRIGGS, F. N. & WISE, R. M. (1974) Effects of hypertonicity on resting and contracting frog skeletal muscles.Am. J. Physiol. 226, 855–63.Google Scholar
  17. JEWELL, B. R. & RUEGG, J. C. (1966) Oscillatory contraction of insect flight muscle after glycerol extraction.Proc. R. Soc. Lond., Ser. B 164, 428–59.Google Scholar
  18. KERRICK, W. G. L. & DONALDSON, S. K. B. (1972) The effects of Mg2+ on submaximum Ca2+-activated tension in skinned fibres of frog skeletal muscle.Biochem. biophys. Acta 275, 117–22.Google Scholar
  19. KRETSINGER, R. J. (1980) Structure and evolution of calcium-modulated proteins.CRC Crit. Rev. Biochem. 8, 119–74.Google Scholar
  20. LEHMAN, W. (1978) Thick-filament-linked calcium regulation in vertebrate striated muscle.Nature 274, 80–1.Google Scholar
  21. LEHMAN, W. & SZENT-GYÖRGYI, A. G. (1975) Regulation of muscular contraction. Distribution of actin control and myosin control in the Animal Kingdom.J. gen. Physiol. 66, 1–30.Google Scholar
  22. LEVINE, B. A., THORNTON, J. M., FERNANDES, R., KELLY, C. M. & MERCOLA, D. (1978) Comparison of the calcium and magnesium-induced structural changes of troponin-C.Biochem. biophys. Acta 535, 11–24.Google Scholar
  23. LONG, C. (1961)Biochemist's Handbook. London: E. & F. N. Spon.Google Scholar
  24. LOXDALE, H. D. (1976) A method for the continuous assay of pico-mole quantities of ADP released from glycerol-extracted skeletal muscle fibres on MgATP activation.J. Physiol. 260, 4–5P.Google Scholar
  25. LOXDALE, H. D. (1980)Molecular parameters of diverse muscle systems. D.Phil. thesis, University of Oxford.Google Scholar
  26. MARSTON, S. B. & WEBER, A. M. (1975) The dissociation constant of the actin-heavy meromyosin subfragment-1 complex.Biochemistry 14, 3868–73.Google Scholar
  27. McLACHLAN, A. D. & STEWART, M. (1976) The 14-fold periodicity in α-tropomyosin and the interaction with actin.J. molec. Biol. 103, 271–98.Google Scholar
  28. NANNINGA, L. B. (1961) Calculation of free magnesium, calcium and potassium in muscle.Biochem. biophys. Acta 54, 338–44.Google Scholar
  29. NANNINGA, L. B. & KEMPEN, R. (1971) Role of magnesium and calcium in the first and second contraction of glycerin-extracted muscle fibres.Biochemistry 10, 2449–56.Google Scholar
  30. PERRIN, D. D. & SAYCE, I. G. (1967) Computer calculation of equilibrium concentrations in mixtures of metal ions and complexing species.Talanta 14, 833–42.Google Scholar
  31. REUBEN, J. P., BRANDT, P. W., BERMAN, M. & GRUNDFEST, H. (1971) Regulation of tension in the skinned crayfish muscle fibre. I. Contraction and relaxation in the absence of Ca (pCa>9).J. gen. Physiol. 57, 385–408.Google Scholar
  32. SACKTOR, B. (1953) Investigations on the mitochondria of the housefly,Musca domestica L.J. gen. Physiol. 36, 371–87.Google Scholar
  33. SEAMONS, K. B., HARTSHORNE, D. J. & BOTHNER-BY, A. A. (1977) Ca2+ and Mg2+-dependent conformations of troponin-C as determined by1H and19F nuclear magnetic resonance.Biochemistry 16, 4039–46.Google Scholar
  34. SILLEN, L. G. & MARTELL, A. E. (1971) Stability constants of metal-ion complexes.Chem. Soc. Sp. Publ. 25, Suppl. 1.Google Scholar
  35. STEIGER, G. J. (1977) Stretch activation and tension transients in cardiac, skeletal and insect flight muscle. InInsect Flight Muscle (edited by TREGEAR, R. T.), pp. 221–268. Amsterdam: North-Holland.Google Scholar
  36. THAMES, M. D., TEICHHOLZ, L. E. & PODOLSKY, R. J. (1974) Ionic strength and the contraction kinetics of skinned muscle fibres.J. gen. Physiol. 63, 509–30.Google Scholar
  37. VOM BROCKE, H. H. (1966) The activating effects of Ca2+ on the contractile system of insect fibrillar flight muscle.Pflügers Arch. 290, 70–9.Google Scholar
  38. WALLIMANN, T. & SZENT-GYÖRGYI, A. G. (1981) An immunological approach to myosin light-chain function in thick filament linked regulation: effects of anti-scallop myosin light chain antibodies. Possible regulatory role for the essential light chain.Biochemistry 20, 1188–97.Google Scholar
  39. WILKINSON, G. N. (1961) Statistical estimation in enzyme kinetics.Biochem. J. 80, 324–32.Google Scholar
  40. WOLEDGE, R. C. (1968) The energetics of tortoise muscle.J. Physiol. 197, 685–707.Google Scholar
  41. YANAGIDA, T., KURANAGA, I. & INOUE, A. (1982) Interaction of myosin with thin filaments during contraction and relaxation: effect of ionic strength.J. Biochem. 92, 407–12.Google Scholar

Copyright information

© Chapman and Hall Ltd 1983

Authors and Affiliations

  • H. D. Loxdale
    • 1
  • R. T. Tregear
    • 2
  1. 1.Zoology DepartmentUniversity of OxfordOxfordUK
  2. 2.ARC Institute of Animal PhysiologyCambridgeUK

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