Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1981 Apr 1;195(1):317–327. doi: 10.1042/bj1950317

Electrophoretic analysis of proteins from single bovine muscle fibres.

O A Young, C L Davey
PMCID: PMC1162888  PMID: 6458285

Abstract

A number of single fibres were isolated by dissection of four bovine masseter (ma) muscles, three rectus abdominis (ra) muscles and eight sternomandibularis (sm) muscles. By histochemical criteria these muscles contain respectively, solely slow fibres (often called type I), predominantly fast fibres (type II), and a mixture of fast and slow. The fibres were analysed by conventional sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and the gels stained with Coomassie Blue. Irrespective of the muscle, every fibre could be classed into one of two broad groups based on the mobility of proteins in the range 135000-170000 daltons. When zones containing myosin heavy chain were cut from the single-fibre gel tracks and 'mapped' [Cleveland, Fischer, Kirschner & Laemmli (1977) J. Biol. Chem. 252, 1102-1106] with Staphylococcus proteinase, it was found that one group always contained fast myosin heavy chain, whereas the second group always contained the slow form. Moreover, a relatively fast-migrating alpha-tropomyosin was associated with the fast myosin group and a slow-migrating form with the slow myosin group. All fibres also contained beta-tropomyosin; the coexistence of alpha- and beta-tropomyosin is at variance with evidence that alpha-tropomyosin is restricted to fast fibres [Dhoot & Perry (1979) Nature (London) 278, 714-718]. Fast fibres containing the expected fast light chains and troponins I and C fast were identified in the three ra muscles, but in only four sm muscles. In three other sm muscles, all the fast fibres contained two troponins I and an additional myosin light chain that was more typical of myosin light chain 1 slow. The remaining sm muscle contained a fast fibre type that was similar to the first type, except that its myosin light chain 1 was more typical of the slow polymorph. Troponin T was bimorphic in all fast fibres from a ra muscles and in at least some fast fibres from one sm muscle. Peptide 'mapping' revealed two forms of fast myosin heavy chain distributed among fast fibres. Each form was associated with certain other proteins. Slow myosin heavy chain was unvarying in three slow fibre types identified. Troponin I polymorphs were the principal indicator of slow fibre types. The myofibrillar polymorphs identified presumably contribute to contraction properties, but beyond cud chewing involving ma muscle, nothing is known of the conditions that gave rise to the variable fibre composites in sm and ra muscles.

Full text

PDF
317

Images in this article

320Fig. 1.
on p.320
320Fig. 2.
on p.320
321Fig. 3.
on p.321
322Fig. 4.
on p.322
324Fig. 5.
on p.324
325Fig. 6.
on p.325

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amphlett G. W., Perry S. V., Syska H., Brown M. D., Vrbova G. Cross innervation and the regulatory protein system of rabbit soleus muscle. Nature. 1975 Oct 16;257(5527):602–604. doi: 10.1038/257602a0. [DOI] [PubMed] [Google Scholar]
  2. BAILEY K. End-group assay in some proteins of the keratin-myosin group. Biochem J. 1951 Jun;49(1):23–27. doi: 10.1042/bj0490023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brooke M. H., Kaiser K. K. Muscle fiber types: how many and what kind? Arch Neurol. 1970 Oct;23(4):369–379. doi: 10.1001/archneur.1970.00480280083010. [DOI] [PubMed] [Google Scholar]
  4. Brooke M. H., Kaiser K. K. Trophic functions of the neuron. II. Denervation and regulation of muscle. The use and abuse of muscle histochemistry. Ann N Y Acad Sci. 1974 Mar 22;228(0):121–144. doi: 10.1111/j.1749-6632.1974.tb20506.x. [DOI] [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Cummins P., Perry S. V. The subunits and biological activity of polymorphic forms of tropomyosin. Biochem J. 1973 Aug;133(4):765–777. doi: 10.1042/bj1330765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dhoot G. K., Frearson N., Perry S. V. Polymorphic forms of troponin T and troponin C and their localization in striated muscle cell types. Exp Cell Res. 1979 Sep;122(2):339–350. doi: 10.1016/0014-4827(79)90310-0. [DOI] [PubMed] [Google Scholar]
  8. Dhoot G. K., Perry S. V. Distribution of polymorphic forms of troponin components and tropomyosin in skeletal muscle. Nature. 1979 Apr 19;278(5706):714–718. doi: 10.1038/278714a0. [DOI] [PubMed] [Google Scholar]
  9. Ebashi S., Wakabayashi T., Ebashi F. Troponin and its components. J Biochem. 1971 Feb;69(2):441–445. doi: 10.1093/oxfordjournals.jbchem.a129486. [DOI] [PubMed] [Google Scholar]
  10. Etlinger J. D., Zak R., Fischman D. A. Compositional studies of myofibrils from rabbit striated muscle. J Cell Biol. 1976 Jan;68(1):123–141. doi: 10.1083/jcb.68.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gauthier G. F., Lowey S. Distribution of myosin isoenzymes among skeletal muscle fiber types. J Cell Biol. 1979 Apr;81(1):10–25. doi: 10.1083/jcb.81.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hoh J. F. Light chain distribution of chicken skeletal muscle myosin isoenzymes. FEBS Lett. 1978 Jun 15;90(2):297–300. doi: 10.1016/0014-5793(78)80390-1. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Leet N. G., Locker R. H. A prolonged pre-rigor condition in aerobic surfaces of ox muscle. J Sci Food Agric. 1973 Oct;24(10):1181–1191. doi: 10.1002/jsfa.2740241007. [DOI] [PubMed] [Google Scholar]
  15. Lowey S., Risby D. Light chains from fast and slow muscle myosins. Nature. 1971 Nov 12;234(5324):81–85. doi: 10.1038/234081a0. [DOI] [PubMed] [Google Scholar]
  16. Lowry C. V., Kimmey J. S., Felder S., Chi M. M., Kaiser K. K., Passonneau P. N., Kirk K. A., Lowry O. H. Enzyme patterns in single human muscle fibers. J Biol Chem. 1978 Nov 25;253(22):8269–8277. [PubMed] [Google Scholar]
  17. Lutz H., Weber H., Billeter R., Jenny E. Fast and slow myosin within single skeletal muscle fibres of adult rabbits. Nature. 1979 Sep 13;281(5727):142–144. doi: 10.1038/281142a0. [DOI] [PubMed] [Google Scholar]
  18. Offer G., Moos C., Starr R. A new protein of the thick filaments of vertebrate skeletal myofibrils. Extractions, purification and characterization. J Mol Biol. 1973 Mar 15;74(4):653–676. doi: 10.1016/0022-2836(73)90055-7. [DOI] [PubMed] [Google Scholar]
  19. PADYKULA H. A., HERMAN E. The specificity of the histochemical method for adenosine triphosphatase. J Histochem Cytochem. 1955 May;3(3):170–195. doi: 10.1177/3.3.170. [DOI] [PubMed] [Google Scholar]
  20. Peter J. B., Barnard R. J., Edgerton V. R., Gillespie C. A., Stempel K. E. Metabolic profiles of three fiber types of skeletal muscle in guinea pigs and rabbits. Biochemistry. 1972 Jul 4;11(14):2627–2633. doi: 10.1021/bi00764a013. [DOI] [PubMed] [Google Scholar]
  21. Pette D., Schnez U. Coexistence of fast and slow type myosin light chains in single muscle fibres during transformation as induced by long term stimulation. FEBS Lett. 1977 Nov 1;83(1):128–130. doi: 10.1016/0014-5793(77)80656-x. [DOI] [PubMed] [Google Scholar]
  22. Pette D., Schnez U. Myosin light chain patterns of individual fast and slow-twitch fibres of rabbit muscles. Histochemistry. 1977 Oct 22;54(2):97–107. doi: 10.1007/BF00489668. [DOI] [PubMed] [Google Scholar]
  23. Pope B. J., Wagner P. D., Weeds A. G. Heterogeneity of myosin heavy chains in subfragment-1 isoenzymes rabbit skeletal myosin. J Mol Biol. 1977 Jan 25;109(3):470–473. doi: 10.1016/s0022-2836(77)80024-7. [DOI] [PubMed] [Google Scholar]
  24. Pope B. J., Wagner P. D., Weeds A. G. Heterogeneity of myosin heavy chains in subfragment-1 isoenzymes rabbit skeletal myosin. J Mol Biol. 1977 Jan 25;109(3):470–473. doi: 10.1016/s0022-2836(77)80024-7. [DOI] [PubMed] [Google Scholar]
  25. Scopes R. K., Penny I. F. Subunit sizes of muscle proteins, as determined by sodium dodecyl sulphate gel electrophoresis. Biochim Biophys Acta. 1971 May 25;236(2):409–415. doi: 10.1016/0005-2795(71)90221-2. [DOI] [PubMed] [Google Scholar]
  26. Starr R., Offer G. Polarity of the myosin molecule. J Mol Biol. 1973 Nov 25;81(1):17–31. doi: 10.1016/0022-2836(73)90244-1. [DOI] [PubMed] [Google Scholar]
  27. Trinick J., Lowey S. M-protein from chicken pectoralis muscle: isolation and characterization. J Mol Biol. 1977 Jun 25;113(2):343–368. doi: 10.1016/0022-2836(77)90146-2. [DOI] [PubMed] [Google Scholar]
  28. Weeds A. G., Hall R., Spurway N. C. Characterization of myosin light chains from histochemically identified fibres of rabbit psoas muscle. FEBS Lett. 1975 Jan 1;49(3):320–324. doi: 10.1016/0014-5793(75)80776-9. [DOI] [PubMed] [Google Scholar]
  29. d'Albis A., Pantaloni C., Bechet J. J. Structural relationship of myosin isoenyzmes: proteolytic digestion patterns of heavy chain components from fast muscles, and comparison with other muscle types. FEBS Lett. 1979 Oct 1;106(1):81–84. doi: 10.1016/0014-5793(79)80699-7. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES