Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Oct;87(19):7453–7456. doi: 10.1073/pnas.87.19.7453

Contraction of myofibrils in the presence of antibodies to myosin subfragment 2.

W F Harrington 1, T Karr 1, W B Busa 1, S J Lovell 1
PMCID: PMC54765  PMID: 2217176

Abstract

In a muscle-based version of in vitro motility assays, the unloaded shortening velocity of rabbit skeletal myofibrils has been determined in the presence and absence of affinity-column-purified polyclonal antibodies directed against the subfragment-2 region of myosin. Contraction was initiated by photohydrolysis of caged ATP and the time dependence of shortening was monitored by an inverted microscope equipped with a video camera. Antibody-treated myofibrils undergo unloaded shortening in a fast phase with initial rates and half-times comparable to control (untreated) myofibrils, despite a marked reduction in the isometric force of skinned muscle fibers in the presence of the antibodies. In antibody-treated myofibrils, this process is followed by a much slower phase of contraction, terminating in elongated structures with well-defined sarcomere spacings (approximately 1 micron) in contrast to the supercontracted globular state of control myofibrils. These results suggest that although the unloaded shortening of myofibrils (and in vitro motility of actin filaments over immobilized myosin heads) can be powered by myosin heads, the subfragment-2 region as well as the myosin head contributes to force production in actively contracting muscle.

Full text

PDF
7453

Images in this article

7454Image
on p.7454
7455Image
on p.7455
7455Image
on p.7455

Selected References

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

  1. Barnett V. A., Thomas D. D. Microsecond rotational motion of spin-labeled myosin heads during isometric muscle contraction. Saturation transfer electron paramagnetic resonance. Biophys J. 1989 Sep;56(3):517–523. doi: 10.1016/S0006-3495(89)82698-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Collier V. L., Kronert W. A., O'Donnell P. T., Edwards K. A., Bernstein S. I. Alternative myosin hinge regions are utilized in a tissue-specific fashion that correlates with muscle contraction speed. Genes Dev. 1990 Jun;4(6):885–895. doi: 10.1101/gad.4.6.885. [DOI] [PubMed] [Google Scholar]
  3. Cooke R., Crowder M. S., Thomas D. D. Orientation of spin labels attached to cross-bridges in contracting muscle fibres. Nature. 1982 Dec 23;300(5894):776–778. doi: 10.1038/300776a0. [DOI] [PubMed] [Google Scholar]
  4. Duong A. M., Reisler E. Binding of myosin to actin in myofibrils during ATP hydrolysis. Biochemistry. 1989 Feb 7;28(3):1307–1313. doi: 10.1021/bi00429a054. [DOI] [PubMed] [Google Scholar]
  5. Fajer P. G., Fajer E. A., Thomas D. D. Myosin heads have a broad orientational distribution during isometric muscle contraction: time-resolved EPR studies using caged ATP. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5538–5542. doi: 10.1073/pnas.87.14.5538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. George E. L., Ober M. B., Emerson C. P., Jr Functional domains of the Drosophila melanogaster muscle myosin heavy-chain gene are encoded by alternatively spliced exons. Mol Cell Biol. 1989 Jul;9(7):2957–2974. doi: 10.1128/mcb.9.7.2957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harrington W. F. A mechanochemical mechanism for muscle contraction. Proc Natl Acad Sci U S A. 1971 Mar;68(3):685–689. doi: 10.1073/pnas.68.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Harrington W. F. On the origin of the contractile force in skeletal muscle. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5066–5070. doi: 10.1073/pnas.76.10.5066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Huxley A. F., Simmons R. M. Proposed mechanism of force generation in striated muscle. Nature. 1971 Oct 22;233(5321):533–538. doi: 10.1038/233533a0. [DOI] [PubMed] [Google Scholar]
  10. Huxley H. E., Kress M. Crossbridge behaviour during muscle contraction. J Muscle Res Cell Motil. 1985 Apr;6(2):153–161. doi: 10.1007/BF00713057. [DOI] [PubMed] [Google Scholar]
  11. Huxley H. E. Sliding filaments and molecular motile systems. J Biol Chem. 1990 May 25;265(15):8347–8350. [PubMed] [Google Scholar]
  12. Huxley H. E. The mechanism of muscular contraction. Science. 1969 Jun 20;164(3886):1356–1365. doi: 10.1126/science.164.3886.1356. [DOI] [PubMed] [Google Scholar]
  13. Hynes T. R., Block S. M., White B. T., Spudich J. A. Movement of myosin fragments in vitro: domains involved in force production. Cell. 1987 Mar 27;48(6):953–963. doi: 10.1016/0092-8674(87)90704-5. [DOI] [PubMed] [Google Scholar]
  14. Kishino A., Yanagida T. Force measurements by micromanipulation of a single actin filament by glass needles. Nature. 1988 Jul 7;334(6177):74–76. doi: 10.1038/334074a0. [DOI] [PubMed] [Google Scholar]
  15. Lovell S., Karr T., Harrington W. F. Suppression of contractile force in muscle fibers by antibody to myosin subfragment 2. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1849–1853. doi: 10.1073/pnas.85.6.1849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mahdavi V., Chambers A. P., Nadal-Ginard B. Cardiac alpha- and beta-myosin heavy chain genes are organized in tandem. Proc Natl Acad Sci U S A. 1984 May;81(9):2626–2630. doi: 10.1073/pnas.81.9.2626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McNally E. M., Kraft R., Bravo-Zehnder M., Taylor D. A., Leinwand L. A. Full-length rat alpha and beta cardiac myosin heavy chain sequences. Comparisons suggest a molecular basis for functional differences. J Mol Biol. 1989 Dec 5;210(3):665–671. doi: 10.1016/0022-2836(89)90141-1. [DOI] [PubMed] [Google Scholar]
  18. Morano I., Arndt H., Gärtner C., Rüegg J. C. Skinned fibers of human atrium and ventricle: myosin isoenzymes and contractility. Circ Res. 1988 Mar;62(3):632–639. doi: 10.1161/01.res.62.3.632. [DOI] [PubMed] [Google Scholar]
  19. Moss R. L., Giulian G. G., Greaser M. L. Effects of EDTA treatment upon the protein subunit composition and mechanical properties of mammalian single skeletal muscle fibers. J Cell Biol. 1983 Apr;96(4):970–978. doi: 10.1083/jcb.96.4.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pope B., Hoh J. F., Weeds A. The ATPase activities of rat cardiac myosin isoenzymes. FEBS Lett. 1980 Sep 8;118(2):205–208. doi: 10.1016/0014-5793(80)80219-5. [DOI] [PubMed] [Google Scholar]
  21. Schwartz K., Lecarpentier Y., Martin J. L., Lompré A. M., Mercadier J. J., Swynghedauw B. Myosin isoenzymic distribution correlates with speed of myocardial contraction. J Mol Cell Cardiol. 1981 Dec;13(12):1071–1075. doi: 10.1016/0022-2828(81)90297-2. [DOI] [PubMed] [Google Scholar]
  22. Somlyo A. V., Goldman Y. E., Fujimori T., Bond M., Trentham D. R., Somlyo A. P. Cross-bridge kinetics, cooperativity, and negatively strained cross-bridges in vertebrate smooth muscle. A laser-flash photolysis study. J Gen Physiol. 1988 Feb;91(2):165–192. doi: 10.1085/jgp.91.2.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Toyoshima Y. Y., Kron S. J., McNally E. M., Niebling K. R., Toyoshima C., Spudich J. A. Myosin subfragment-1 is sufficient to move actin filaments in vitro. Nature. 1987 Aug 6;328(6130):536–539. doi: 10.1038/328536a0. [DOI] [PubMed] [Google Scholar]
  24. Ueno H., Harrington W. F. An enzyme-probe study of motile domains in the subfragment-2 region of myosin. J Mol Biol. 1984 Dec 15;180(3):667–701. doi: 10.1016/0022-2836(84)90032-9. [DOI] [PubMed] [Google Scholar]
  25. Ueno H., Harrington W. F. Cross-linking within the thick filaments of muscle and its effect on contractile force. Biochemistry. 1987 Jun 16;26(12):3589–3596. doi: 10.1021/bi00386a051. [DOI] [PubMed] [Google Scholar]
  26. Ueno H., Harrington W. F. Local melting in the subfragment-2 region of myosin in activated muscle and its correlation with contractile force. J Mol Biol. 1986 Jul 5;190(1):69–82. doi: 10.1016/0022-2836(86)90076-8. [DOI] [PubMed] [Google Scholar]
  27. Ueno H., Harrington W. F. Temperature-dependence of local melting in the myosin subfragment-2 region of the rigor cross-bridge. J Mol Biol. 1986 Jul 5;190(1):59–68. doi: 10.1016/0022-2836(86)90075-6. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES