Skip to main content
SpringerLink
Log in

Stiffness of carbodiimide-crosslinked glycerinated muscle fibres in rigor and relaxing solutions at high salt concentrations

  • Papers
  • Published:
Journal of Muscle Research & Cell Motility Aims and scope Submit manuscript

Summary

In this article, we have applied a crosslinking technique with a water-soluble carbodiimide to single glycerol-extracted muscle fibres from the rabbit. We have measured the stiffness of the fibres in a relaxing solution at high salt concentration. These fibres were crosslinked to varying extents in the rigor state. The relaxing solution caused uncrosslinked crossbridge heads (S1) to detach. High salt concentrations were used because the fibres were not activated by the crosslinked crossbridges under these conditions, although they were at physiological ionic strength. We found (1) a linear correlation between the extent of S1 crosslinking to thin filaments and the stiffness and (2) that the stiffness in the relaxing solution of muscle fibres with all the S1 heads crosslinked to thin filaments was the same as the rigor stiffness of the fibres before crosslinking. We conclude that the sarcomere compliance is mostly a property of the crossbridges (with more than 65% of the crossbridge compliance in the S1 portions and less than 35% in the S2 portion) and little of other sarcomere structures. In an earlier paper [Kimura & Tawada,Biophys. J. 603–10 (1984)], we demonstrated that the S2 portion of the crossbridge was stiff. It then follows that the crossbridge compliance, and thus the sarcomere compliance, is a property of the S1 heads. Assuming that the S1 portion of the crossbridges in rigor strained muscle fibres is bent, we calculated the Young's modulus of the S1 portion and found that it is about 102 MN m−2. Because this order of magnitude is reasonable in terms of globular protein elasticity, bending is likely to be the nature of the S1 compliance in rigor muscle fibres.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Botts, J., Takashi, R., Torgerson, P., Hozumi, T., Muhlard, A., Mornet, D. &Morales, M. F. (1984) On the mechanism of energy transduction in myosin subfragment 1.Proc. natn. Acad. Sci. U.S.A. 81, 2060–4.

    Google Scholar 

  • Cooke, R. (1972) A new method for producing myosin subfragment 1.Biochem. Biophys. Res. Commun. 49, 1021–8.

    Google Scholar 

  • Cooke, R. (1981) Stress does not alter the conformation of a domain of the myosin cross-bridge in rigor muscle fibres.Nature, Lond. 294, 570–1.

    Google Scholar 

  • Craig, R., Szent-Györgyi, A. G., Beese, L., Flicker, P., Vibert, P. &Cohen, C. (1980) Electron microscopy of thin filaments decorated with a Ca2+-regulated myosin.J. molec. Biol. 140, 35–55.

    Google Scholar 

  • Cross, R. A., Bardsley, R. G., Ledward, D. A., Small, J. V. &Sobieszek, A. (1984) Conformational stability of the myosin rod.Eur. J. Biochem. 145, 305–10.

    Google Scholar 

  • Eisenberg, E. &Hill, T. L. (1978) A cross-bridge model of muscle contraction.Prog. Biophys. molec. Biol. 33, 55–82.

    Google Scholar 

  • Eisenberg, E. &Hill, T. L. (1985) Muscle contraction and free energy transduction in biological systems.Science, N.Y. 227, 999–1006.

    Google Scholar 

  • Elliott, A. &Offer, G. (1978) Shape and flexibility of the myosin molecule.J. molec. Biol. 123, 505–19.

    Google Scholar 

  • Ford, L. E., Huxley, A. F. &Simmons, R. M. (1977) Tension responses to sudden length change in stimulated frog muscle fibres near slack length.J. Physiol., Lond. 269, 441–515.

    Google Scholar 

  • Ford, L. E., Huxley, A. F. &Simmons, R. M. (1981) The relation between stiffness and filament overlap in stimulated frog muscle fibres.J. Physiol., Lond. 311, 219–49.

    Google Scholar 

  • Goldman, Y. E. &Simmons, R. M. (1977) Active and rigor muscle stiffness.J. Physiol., Lond. 269, 55–7p.

    Google Scholar 

  • Güth, K. &Kuhn, H. J. (1978) Stiffness and tension during and after sudden length changes of glycerinated rabbit psoas fibers.Biophys. Struct. Mech. 4, 223–36.

    Google Scholar 

  • Harrington, W. F. (1971) A mechanochemical mechanism for muscle contraction.Proc. natn. Acad. Sci. U.S.A. 68, 685–9.

    Google Scholar 

  • Harrington, W. F. (1979) On the origin of the contractile force in skeletal muscle.Proc. natn. Acad. Sci. U.S.A. 76, 5066–70.

    Google Scholar 

  • Heaphy, S. &Tregear, R. (1984) Stoichiometry of covalent actin-subfragment 1 complexes formed on reaction with a zero-length cross-linking compound.Biochemistry 23, 2211–14.

    Google Scholar 

  • Heuser, J. E. &Cooke, R. (1983) Actin-myosin interactions visualized by the quick-freeze, deep-etch replica technique.J. molec. Biol. 169, 97–122.

    Google Scholar 

  • Highsmith, S., Akasaka, K., Konrad, M., Goody, R., Holmes, K., Wade-Jardetzky, N. &Jardetzky, O. (1979) Internal motions in myosin.Biochemistry 18, 4238–44.

    Google Scholar 

  • Huxley, A. F. (1957) Muscle structure and theories of contraction.Prog. Biophys. biophys. Chem. 7, 255–318.

    Google Scholar 

  • Huxley, A. F. (1974) Review lecture on muscular contraction.J. Physiol., Lond. 243, 1–43.

    Google Scholar 

  • Huxley, A. F. &Simmons, R. M. (1971) Proposed mechanism of force generation in striated muscle.Nature, Lond. 233, 533–8.

    Google Scholar 

  • Huxley, H. E., Simmons, R. M., Faruqi, A. R., Kress, M., Bordas, J. &Koch, M. H. J. (1983) Changes in the X-ray reflections from contracting muscle during rapid mechanical transients and their structural implications.J. molec. Biol. 169, 469–506,

    Google Scholar 

  • Kimura, M. &Tawada, K. (1984) Is the SII portion of the cross-bridge in glycerinated rabbit psoas fibers compliant in the rigor state?Biophys. J. 45, 603–10.

    Google Scholar 

  • Knight, P. &Trinick, J. (1984) Structure of the myosin projections on native thick filaments from vertebrate skeletal muscle.J. molec. Biol. 177, 461–82.

    Google Scholar 

  • Love, A. E. H. (1934)A Treatise on the Mathematical Theory of Elasticity, P. 329. London: Cambridge University Press.

    Google Scholar 

  • Matsubara, I., Goldman, Y. E. &Simmons, R. M. (1984) Changes in the lateral filament spacing of skinned muscle fibers when cross-bridges attach.J. molec. Biol. 173, 15–33.

    Google Scholar 

  • McCammon, J. A. (1984) Protein dynamics.Rep. Prog. Phys. 47, 1–46.

    Google Scholar 

  • Mendelson, R. A. &Wagner, P. D. (1984) X-ray scattering by single-headed heavy meromyosin: Cleavage of the myosin head from the rod does not change its shape.J. molec. Biol. 177, 153–71.

    Google Scholar 

  • Mornet, D., Bertrand, R., Pantel, P., Audemard, E. &Kassab, R. (1981) Structure of the actin-myosin interface.Nature, Lond. 292, 301–6.

    Google Scholar 

  • Naylor, G. R. S. &Podolsky, R. J. (1981) X-ray diffraction of strained muscle fibers in rigor.Proc. natn. Acad. Sci. U.S.A. 78, 5559–63.

    Google Scholar 

  • Perry, S. V. (1955) Myosin adenosine triphosphatase.Meth. Enzym. 2, 582–8.

    Google Scholar 

  • Prince, H. P., Trayer, H. R., Henry, G. D., Trayer, I. P., Dalgarno, D. C., Levine, B. A., Cary, P. D. &Turner, C. (1981) Proton nuclear-magnetic-resonance spectroscopy of myosin subfragment 1 isozymes.Eur. J. Biochem. 121, 213–9.

    Google Scholar 

  • Suezaki, Y. &Gō, N. (1976) Fluctuations and mechanical strength of α-helices of polyglycine and poly(L-alanine).Biopolymers 15, 2137–53.

    Google Scholar 

  • Sutoh, K. (1983) Mapping of actin-binding sites on heavy chain of myosin subfragment 1.Biochemistry 22, 1579–85.

    Google Scholar 

  • Sutoh, K., Yamamoto, K. &Wakabayashi, T. (1984) Electron microscopic visualization of the SH1 thiol of myosin by the use of an avidin-biotin system.J. molec. Biol. 178, 323–39.

    Google Scholar 

  • Suzuki, S. &Sugi, H. (1983) Extensibility of the myofilaments in vertebrate skeletal muscle as revealed by stretching rigor muscle fibers.J. gen. Physiol. 81, 531–46.

    Google Scholar 

  • Takahashi, K. (1978) Topography of the myosin molecule as visualized by an improved negative staining method.J. Biochem., Tokyo 83, 905–8.

    Google Scholar 

  • Tanaka, H., Sugi, H. &Hashizume, H. (1984) Effect of stretch on the equatorial X-ray diffraction pattern from frog skeletal muscle in rigor. InContractile Mechanism in Muscle (edited byPollack, G. H. andSugi, H.), pp. 203–5. New York: Plenum Press.

    Google Scholar 

  • Tawada, K. &Kimura, M. (1984a) Cross-linking studies related to the location of the rigor compliance in glycerinated rabbit psoas fibers: Is the SII portion of the cross-bridge compliant? InContractile Mechanisms in Muscle (edited byPollack, G. H. andSugi, H.), pp. 385–96. New York: Plenum Press.

    Google Scholar 

  • Tawada, K. &Kimura, M. (1984b) Stiffness of glycerinated rabbit psoas fibers in rigor: Filament-overlap relation.Biophys. J. 45, 593–602.

    Google Scholar 

  • Toyoshima, C. &Wakabayashi, T. (1985) Threedimensional image analysis of the complex of thin filaments and myosin molecules from skeletal muscle. IV. Reconstruction from minimal- and high-dose images of the actin-tropomyosin-myosin subfragment-1 complex.J. Biochem., Tokyo 97, 219–43.

    Google Scholar 

  • Tsong, T. Y., Karr T. &Harrington, W. F. (1979) Rapid helix-coil transitions in the S-2 region of myosin.Proc. natn. Acad. Sci. U.S.A. 76, 1109–13.

    Google Scholar 

  • Ueno, H., Rodgers, M. E. &Harrington, W. F. (1983) Self-association of a high molecular weight subfragment-2 of myosin induced by divalent metal ions.J. molec. Biol. 168, 207–28.

    Google Scholar 

  • Wagner, P. D. &Weeds, A. G. (1977) Studies on the role of myosin alkali light chains.J. molec. Biol. 109, 455–73.

    Google Scholar 

  • Weber, A. &Murray, J. M. (1973) Molecular control mechanisms in muscle contraction.Physiol. Rev. 53, 612–73.

    Google Scholar 

  • Weber, K. &Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfatepolyacrylamide gel electrophoresis.J. biol. Chem. 244, 4406–12.

    Google Scholar 

  • Weeds, A. G. &Taylor, R. S. (1975) Separation of subfragment-1 isozymes from rabbit skeletal muscle myosin.Nature, Lond. 257, 54–6.

    Google Scholar 

  • Winkelmann, D. A., Mekeel, H. &Rayment, I. (1985) Packing analysis of crystalline myosin subfragment-1: Implications for the size and shape of the myosin head.J. molec. Biol. 181, 487–501.

    Google Scholar 

  • Yamamoto, K., Sekine, T. &Sutoh, K. (1984) Spatial relationship between SH1 and the actin binding site on myosin subfragment-1 surface.FEBS Lett. 176, 75–8.

    Google Scholar 

  • Yamamoto, T. &Herzig, J. W. (1978) Series elastic properties of skinned muscle fibers in contraction and rigor.Pflügers Arch. 373, 21–4.

    Google Scholar 

  • Yanagida, T. (1984) Angles of fluorescently labelled myosin heads and actin monomers in contracting and rigor strained muscle fiber. InContractile Mechanisms in Muscle (edited byPollack, G. H. andSugi, H.), pp. 397–411. New York: Plenum Press.

    Google Scholar 

  • Young, D. M., Himmelfarb, S. &Harrington, W. F. (1965) On the structural assembly of the polypeptide chains of heavy meromyosin.J. biol. Chem. 240, 2428–36.

    Google Scholar 

Download references

Author information

Author notes

  1. Michio Kimura

    Present address: Department of Basic Research, R & D Laboratories, Hoechst Japan, Minamidai, Kawagome, Japan

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Kyushu University, Fukuoka, 812, Fukuoka, Japan

    Katsuhisa Tawada & Michio Kimura

Authors
  1. Katsuhisa Tawada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michio Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tawada, K., Kimura, M. Stiffness of carbodiimide-crosslinked glycerinated muscle fibres in rigor and relaxing solutions at high salt concentrations. J Muscle Res Cell Motil 7, 339–350 (1986). https://doi.org/10.1007/BF01753655

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01753655

Keywords

Search

Navigation