Summary
The deteriorating effect of μm order of Ca2+ on skinned frog skeletal muscle fibres was studied from the view point of the digestion of proteins by calcium-activated neutral protease (CANP). Tension developed in solutions containing no MgATP (rigor solution) decreased irreversibly with the addition of Ca2+ in quantities of more than 0.1 μm. Low temperature was seen to suppress (Q10>4), and neutral pH to maximize, this decrease in tension. In rigor solution containing Ca2+, SDS electrophoresis indicated that a 95 k dalton component (α-actinin) was released from the fibre; electron micrography showed the disappearance of Z-lines. These results suggest that one of the causes for decrease in rigor tension is the proteolytic activity of CANP, and its inhibitors were shown to be quite useful in experiments on skinned fibre.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bush, W. A., Stromer, M. H., Goll, D. E. &Suzuki, A. (1972) Ca2+-specific removal of Z-lines from rabbit skeletal muscle.J. Cell. Biol. 52, 367–81.
Bodensteiner, J. B. &Engel, A. G. (1978) Intracellular calcium accumulation in Duchenne dystrophy and other myopaties: A study of 567,000 muscle fibers in 114 biopsies.Neurology 28, 439–46.
Cullen, M. J. &Fulthorpe, J. J. (1975) Stages in fibre breakdown in Duchenne muscular dystrophy.J. Neurol. Sci. 24, 179–200.
Ebashi, S. &Endo, M. (1968) Calcium and muscle contraction.Prog. Biophys. Mol. Biol. 18,123–83.
Harafuji, H. &Ogawa, Y. (1980) Re-examination of the apparent binding constant of ethylene glycol bis (β-aminoethyl ether)-N, N, NT, N′-tetraacetic acid with calcium around neutral pH.J. Biochem. 87,1305–12.
Hattori, A. &Takahashi, K. (1982) Calcium-induced weakening of skeletal muscle Z-disks.J. Biochem. 92, 381–90.
Haeberle, J. R., Coolican, S. A., Evan, A. &Hathaway, D. R. (1985) The effect of calcium dependent protease on the ultrastructure and contractile mechanics of skinned uterine smooth muscle.J. Musc. Res. Cell Motil. 6, 347–63.
Higuchi, H. &Umazume, Y. (1985) Localization of the parallel elastic components in frog skinned muscle fibers studied by the dissociation of the A- and I-bands.Biophys. J. 48, 137–47.
Ishiura, S., Murofushi, H., Suzuki, K. &Imahori, K. (1978) Studies of a calcium-activated neutral protease from chicken skeletal muscle.J. Biochem. 84, 225–30.
Kawashima, S., Nomoto, M., Hayashi, M., Inomata, M., Nakamura, M. &Imahori, K. (1984) Comparison of calcium-activated neutral proteases from skeletal muscle of rabbit and chicken.J. Biochem. 95, 95–101.
Maruyama, K., Sawada, H., Kimura, S., Ohashi, K., Higuchi, H. &Umazume, Y. (1984) Connectin filaments in stretched skinned fibers of frog skeletal muscle.J. Cell. Bid. 99, 1391–7.
McGowan, E. B., Shafiq, S. A. &Stracher, A. (1976) Delayed degeneration of dystrophic and normal muscle cell cultures treated with pepstatin, leupeptin, and antipain.Exp. Neurol. 50, 649–57.
Mellgren, R. L. (1980) Canine cardiac calcium-dependent proteases: Resolution of two forms with different requirements for calcium.FEBS Letts. 109, 129–33.
Mellgren, R. L. &Carr, T. C. (1983) The protein inhibitor of calcium-dependent proteases: Purification from bovine heart and possible mechanisms of regulation.Arch. Biochem. Biophys. 225, 779–86.
Natori, R. (1954) The role of myofibrils, sarcoplasma and sarcolemma in muscle-contraction.Jikeikai Med. J. 1, 18–28.
Pearce, G. W. (1966) Electron microscopy in the study of muscular dystrophy.Ann. N.Y. Acad. Sci. 138, 138–50.
Publicover, S. J., Duncan, C. J. &Smith, J. L. (1978) The use of A23187 to demonstrate the role of intracellular calcium in causing ultrastructural damage in mammalian muscle.J. Neuropath. Exp. Neurol. 37, 544–57.
Reddy, M. K., Etlinger, J. D., Rabinowitz, M., Fischman, D. A. &Zak, R. (1975) Removal of Z-lines and α-actinin from isolated myofibrils by a calcium-activated neutral protease.J. Biol. Chem. 250, 4278–84.
Suzuki, S. &Sugi, H. (1983) Extensibility of the myofilaments in vertebrate skeletal muscle as revealed by stretching rigor muscle fibers.J. Gen. Physiol. 81, 531–546.
Umazume, Y., Onodera, S. &Higuchi, H. (1986) Width and lattice spacing in radially compressed frog skinned muscle fibers at various pH values, magnesium ion concentrations and ionic strengths.J. Musc. Res. Cell Motil. 7, 251–8.
Wang, K. (1985) Sarcomere-associated cytoskeletal lattices in striated muscle. InCell and Muscle Motility, Vol. 6. (edited byShay, J. W.) pp. 315–69. Plenum Publishing Corp.
Weber, K. &Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.J. Biol. Chem. 244, 4406–12.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kasuga, N., Umazume, Y. Deterioration induced by physiological concentration of calcium ions in skinned muscle fibres. J Muscle Res Cell Motil 11, 41–47 (1990). https://doi.org/10.1007/BF01833324
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01833324