Journal of Muscle Research & Cell Motility

, Volume 4, Issue 6, pp 717–738 | Cite as

Myosin isozymes in avian skeletal muscles. II. Fractionation of myosin isozymes from adult and embryonic chicken pectoralis muscle by immuno-affinity chromatography

  • Pamela A. Benfield
  • Susan Lowey
  • Denise D. LeBlanc
  • Guillermina S. Waller


Chicken pectoralis consists primarily of large white fibres, which react exclusively with antibodies prepared against adult fast myosin. There is, however, a small region of uniformly red fibres which responds to antibodies against adult slow myosin as well as adult fast myosin. The myosin extracted from this red region is also heterogeneous as shown by the presence of both slow and fast light chains. By means of immunoadsorbents, it has been possible to separate the ‘red myosin’ into a ‘fast’ component and a ‘slow’ component. These two fractions have been characterized with respect to their light and heavy chain content by one-dimensional and two-dimensional gel electrophoresis. The myosin heavy chain was reduced to the smaller fragments required for electrophoresis by proteolytic degradation. We conclude from the electrophoretic patterns that the ‘fast’ and ‘slow’ myosin components from the pectoralis red region closely resemble the myosin from the white region of the pectoralis and the myosin from the slow anterior latissimus dorsi (ALD) muscle.

The demonstration of a ‘slow myosin’ in adult pectoralis muscle raises the possibility that the crossreactivity of embryonic pectoralis myosin with anti-slow (ALD) myosin antibodies might be due to the presence of such slow components in embryonic chicken muscle. Direct isolation of a slow component from embryonic pectoralis was achieved by immunoadsorption, as described for adult mixed muscle myosin. Analysis of the subunit composition by gel electrophoresis shows an enrichment in adult-type slow light chains, but the heavy chain pattern is quite distinct from that of adult slow heavy chain. These studies suggest that several myosin isozymes exist in embryonic chicken pectoralis, but that none is identical to those myosins found in the different fibres of the adult pectoralis muscle.


Heavy Chain Slow Component Pectoralis Muscle Embryonic Chicken Slow Light 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. ARNDT, T. & PEPE, F. A. (1975) Antigenic specificity of red and white muscle myosin.J. Histochem. Cytochem. 23, 159–68.Google Scholar
  2. ATSUMI, S. (1977) Development of neuromuscular junctions of fast and slow muscles in the chick embryo: a light and electron microscopic study.J. Neurocytol. 6, 691–709.Google Scholar
  3. BADER, D., MASAKI, T. & FISCHMAN, D. A. (1982) Immunochemical analysis of myosin heavy chain during avian myogenesisin vivo andin vitro.J. Cell Biol. 95, 763–70.Google Scholar
  4. BANDMAN, E., MATSUDA, R. & STROHMAN, R. C. (1982) Myosin heavy chains from two different adult fast-twitch muscles have different peptide maps but identical mRNAs.Cell 29, 645–50.Google Scholar
  5. BARANY, M. (1967) ATPase activity of myosin correlated with speed of muscle shortening.J. gen. Physiol. 50, 197–218.Google Scholar
  6. BRUGGMANN, S. & JENNY, E. (1975) The immunological specificity of myosins from cross-striated muscles as revealed by quantitative microcomplement fraction and enzyme inhibition by antisera.Biochim. biophys. Acta 412, 39–50.Google Scholar
  7. BURKE, R. E., LEVINE, D. N., TSAIRIS, P. & ZAIAC, F. E., III (1973) Physiological types and histochemical profiles in motor units of the cat gastrocnemius.J. Physiol. 234, 723–48.Google Scholar
  8. BURRIDGE, K. & BRAY, D. (1975) Purification and structural analysis of myosins from brain and other non-muscle tissues.J. molec. Biol. 99, 1–14.Google Scholar
  9. D'ALBIS, A. & GRATZER, W. B. (1973) Electrophoretic examination of native myosin.FEBS Lett. 97, 330–4.Google Scholar
  10. D'ALBIS, A., PANTALONI, C. & BECHET, J. J. (1979) Structural relationship of myosin isoenzymes. Proteolytic digestion patterns of heavy chain components from fast muscles, and comparison with other muscles types.FEBS Lett. 106, 81–4.Google Scholar
  11. FLINK, I. L., RADER, J. H. & MORKIN, E. (1979) Thyroid hormone stimulates synthesis of a cardiac myosin isozyme. Comparison of the two-dimensional electrophoretic patterns of the cyanogen bromide peptides of cardiac myosin heavy chains from euthyroid and thyrotoxic rabbits.J. biol. Chem. 254, 3105–10.Google Scholar
  12. FRANK, G. & WEEDS, A. G. (1974) The amino acid sequence of the alkali light chains of rabbit skeletal muscle myosin.Eur. J. Biochem. 44, 317–34.Google Scholar
  13. GALLAGHER, M. & ELZINGA, M. (1980) Amino acid sequence of a 21,000 dalton tryptic peptide from myosin.Fed. Proc. 39, 2168.Google Scholar
  14. GAUTHIER, G. F. (1980) Distribution of myosin isoenzymes in adult and developing muscle fibres. InPlasticity of Muscle (edited by PETTE, D.), pp. 83–96. Berlin, New York: de Gruyter.Google Scholar
  15. GAUTHIER, G. F., BURKE, R. E., LOWEY, S. & HOBBS, A. W. (1983) Myosin isozymes in normal and cross-reinnervated cat skeletal muscle fibres.J. Cell Biol. 97, 756–71.Google Scholar
  16. GAUTHIER, G. F. & LOWEY, S. (1977) Polymorphism of myosin among skeletal muscle fibre types.J. Cell Biol. 74, 760–79.Google Scholar
  17. GAUTHIER, G. F. & LOWEY, S. (1979) Distribution of myosin isoenzymes among skeletal muscle fibre types.J. Cell Biol. 81, 10–25.Google Scholar
  18. GAUTHIER, G. F., LOWEY, S., BENFIELD, P. A. & HOBBS, A. W. (1982) Distribution and properties of myosin isozymes in developing avian and mammalian skeletal muscle fibres.J. Cell Biol. 92, 471–84.Google Scholar
  19. GAUTHIER, G. F., LOWEY, S. & HOBBS, A. W. (1978) Fast and slow myosin in developing muscle fibres.Nature 274, 25–9.Google Scholar
  20. GRÖSCHEL-STEWART, U., SCHREIBER, J., MAHLMEISTER, C. & WEBER, K. (1976) Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated muscle myosins.Histochemistry 46, 229–36.Google Scholar
  21. HOH, J. F. Y. (1975) Neural regulation of mammalian fast and slow muscle myosins: An electrophoretic analysis.Biochemistry 14, 742–7.Google Scholar
  22. HOH, J. F. Y., MCGRATH, P. A. & WHITE, R. I. (1976) Electrophoretic analysis of multiple forms of myosin in fast-twitch and slow-twitch muscles of the chick.Biochem. J. 157, 87–95.Google Scholar
  23. HOH, J. F. Y., YEOH, G. P. S., THOMAS, M. A. W. & HIGGINBOTTOM, L. (1979) Structural differences in the heavy chains of rat ventricular myosin isoenzymes.FEBS Lett. 97, 330–4.Google Scholar
  24. HOLT, J. C. & LOWEY, S. (1977) Distribution of alkali light chains in myosin: isolation of isoenzymes.Biochemistry 16, 4398–402.Google Scholar
  25. KELLER, L. R. & EMERSON, C. P., JR (1980) Synthesis of adult myosin light chains by embryonic muscle cultures.Proc. natn. Acad. Sci. 77, 1020–4.Google Scholar
  26. LAEMMLI, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 227, 680–4.Google Scholar
  27. LEGER, J. T., KLOTZ, C., CAVAILLE, F. & MAROTTE, F. (1979) Structural differences between the heavy chains of myosin subfragment-1 from bovine, porcine and human hearts.FEBS Lett. 106, 157–61.Google Scholar
  28. LOWEY, S. (1980) An immunological approach to the isolation of myosin isoenzymes. InPlasticity of Muscle (edited by PETTE, D.), pp. 69–81. Berlin, New York: de Gruyter.Google Scholar
  29. LOWEY, S., BENFIELD, P. A., LeBLANC, D. D. & WALLER, G. S. (1983) Myosin isozymes in avian skeletal muscles. I. Sequential expression of myosin isozymes in developing chicken pectoralis muscles.J. Musc. Res. Cell Motility 4, 695–716.Google Scholar
  30. LOWEY, S., BENFIELD, P. A., LeBLANC, D. D., WALLER, G. S., WINKELMANN, D. A. & GAUTHIER, G. F. (1982) Characterization of myosins from embryonic and developing chicken pectoralis muscle. InMuscle Development: Molecular and Cellular Control (edited by PEARSON, M. L. and EPSTEIN, H. F.), pp. 15–24. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory.Google Scholar
  31. LOWEY, S., BENFIELD, P. A., SILBERSTEIN, L. & LANG, L. M. (1979a) Distribution of light chains in fast skeletal myosin.Nature 282, 522–4.Google Scholar
  32. LOWEY, S. & RISBY, D. (1971) Light chains from fast and slow muscle myosins.Nature 234, 81–5.Google Scholar
  33. LOWEY, S., SILBERSTEIN, L., GAUTHIER, G. F. & HOLT, J. C. (1979b) Isolation and distribution of myosin isoenzymes. InMotility in Cell Function (edited by PEPE, F. A., SANGER, J. W. and NACHMIAS, V. T.), pp. 53–67. New York, London: Academic Press.Google Scholar
  34. MASAKI, T. (1974) Immunochemical comparison of myosins from chicken cardiac, fast white, slow red and smooth muscle.J. Biochem. 76, 441–9.Google Scholar
  35. OAKLEY, B. R., KIRSCH, D. R. & MORRIS, N. R. (1980) A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels.Analyt. Biochem. 105, 361–3.Google Scholar
  36. O'FARRELL, P. H. (1975) High resolution two-dimensional electrophoresis of proteins.J. biol. Chem. 250, 4007–21.Google Scholar
  37. RUSHBROOK, J. I. & STRACHER, A. (1979) Comparison of adult, embryonic, and dystrophic myosin heavy chains from chicken muscle by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and peptide mapping.Proc. natn. Acad. Sci. 76, 4331–4.Google Scholar
  38. SCHACHAT, F., GARCEA, R. L. & EPSTEIN, H. F. (1978) Myosins exist as homodimers of heavy chains: Demonstration with a specific antibody purified by nematode mutant myosin affinity chromatography.Cell 15, 405–11.Google Scholar
  39. SCHACHAT, F. H., HARRIS, H. E. & EPSTEIN, H. F. (1977) Two homogeneous myosins in body wall muscle ofCaenorhabditis elegans.Cell 10, 721–8.Google Scholar
  40. SILBERSTEIN, L. & LOWEY, S. (1981) Isolation and distribution of myosin isoenzymes in chicken pectoralis muscle.J. molec. Biol. 148, 153–89.Google Scholar
  41. SRETER, F. A., BALINT, M. & GERGELY, J. (1975) Structural and functional changes of myosin during development. Comparison with adult fast, slow and cardiac myosin.Devl Biol. 46, 317–25.Google Scholar
  42. STARR, R. & OFFER, G. (1973) Polarity of the myosin molecule.J. molec. Biol. 81, 17–31.Google Scholar
  43. SWITZER, R. C., III, MERRIL, C. R. & SHIFRIN, S. (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels.Analyt. Biochem. 98, 231–7.Google Scholar
  44. WAGNER, P. D. (1977) Fractionation of heavy meromyosin by affinity chromatography.FEBS Lett. 81, 81–5.Google Scholar
  45. WEEDS, A. G. & TAYLOR, R. S. (1975) Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin.Nature 257, 54–6.Google Scholar
  46. WHALEN, R. G., SCHWARTZ, K., BOUVERET, P., SELL, S. M. & GROS, F. (1979) Contractile protein isozymes in muscle development: Identification of an embryonic form of myosin heavy chain.Proc. natn. Acad. Sci. 76, 5197–201.Google Scholar
  47. WINKELMANN, D. A., LOWEY, S. & PRESS, J. L. (1983) Monoclonal antibodies localize changes on myosin heavy chain isozymes during avian myogenesis.Cell 34, 295–306.Google Scholar
  48. ZWEIG, S. E. (1981) The muscle specificity and structure of two closely related fast twitch white muscle myosin heavy chain isozymes.J. biol. Chem. 256, 11847–53.Google Scholar

Copyright information

© Chapman and Hall Ltd 1983

Authors and Affiliations

  • Pamela A. Benfield
    • 1
  • Susan Lowey
    • 1
  • Denise D. LeBlanc
    • 1
  • Guillermina S. Waller
    • 1
  1. 1.Rosenstiel Basic Medical Sciences Research CenterBrandeis UniversityWalthamUSA

Personalised recommendations