Journal of Muscle Research & Cell Motility

, Volume 1, Issue 3, pp 305–320 | Cite as

Evidence for an altered structure of actin-S1 complexes when Mg-adenylylimidodiphosphate binds

  • S. B. Marston
Papers

Summary

Simultaneous measurements of actin-S1 binding and light scatter (or turbidity) were made in the presence and absence of saturating Mg-adenylylimidodiphosphate (Mg.AMP.PNP). It was found that if the actin was covalently labelled on cysteine 363 by dansyl aziridine (DAZ) the ternary complex DAZ actin-S1-Mg.AMP.PNP scattered 36% less light than the binary complex DAZ actin-S1. If the actin was not labelled there was no difference in scatter. The decrease in scatter of DAZ actin-S1 was dependent on Mg.AMP.PNP concentration in a hyperbolic manner with half maximal decrease at 40 µm. No change in light scatter by DAZ actin-S1 was found when H.AMP.PNP or Mg.ADP bound.

The change in light scatter is evidence that the structure of DAZ actin-S1 is altered when Mg.AMP.PNP binds. The possible nature of the structural change and its role in chemo-mechanical transduction is discussed.

Keywords

Cysteine Structural Change Simultaneous Measurement Light Scatter Maximal Decrease 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. ARMITAGE, P. M., TREGEAR, R. T. & MILLER, A. (1975) Effect of activation by calcium on the X-ray diffraction pattern from insect flight muscle.J. molec. Biol. 92, 39–53.Google Scholar
  2. BAGSHAW, C. R., ECCLESTON, J. F., TRENTHAM, D. R., YATES, D. W. & GOODY, R. S. (1972) Transient kinetics of the Mg2+-dependent ATPase of myosin and its proteolytic subfragments.Cold Spring Harb. Symp. quant. Biol. 37, 127–35.Google Scholar
  3. BEINBRECH, G., KUHN, H. J., HERZIG, J. W. & RÜEGG, J. C. (1976) Evidence for two attached myosin crossbridge states of different potential energy.Cytobiologie 12, 385–96.Google Scholar
  4. BURKE, M., REISLER, E. & HARRINGTON, W. F. (1974) Myosin ATP hydrolysis: A mechanism involving a magnesium chelate complex.Proc. natn. Acad. Sci. U.S.A. 70, 3793–6.Google Scholar
  5. CARLSON, F. D. & FRASER, A. B. (1974) Dynamics of F-actin and F-actin complexes.J. molec. Biol. 89, 273–81.Google Scholar
  6. DETMERS, P. A. (1979) Ph.D. thesis. University of Pennsylvania.Google Scholar
  7. ELZINGA, M. & COLLINS, J. H. (1972) The amino acid sequence of rabbit skeletal muscle actin.Cold Spring Harb. Symp. quant. Biol. 37, 1–7.Google Scholar
  8. FINLAYSON, B., LYMN, R. W. & TAYLOR, E. W. (1969) Studies on the kinetics of formation and dissociation of the actomyosin complex.Biochemistry 8, 811–9.Google Scholar
  9. FUJIME, S. & ISHIWATA, S. (1971) Dynamic study of F-actin by quasielastic scattering of laser light.J. molec. Biol. 62, 251–65.Google Scholar
  10. GERGELY, J. & KOHLER, H. (1957) InConference on the Chemistry of Muscle Contraction. Tokyo: Igaku Shoin.Google Scholar
  11. GREENE, L. E. & EISENBERG, E. (1978) The formation of a ternary complex: actin, adenylylimidodiphosphate, and the subfragments of myosin.Proc. natn. Acad. Sci. U.S.A. 75, 54–8.Google Scholar
  12. GREENE, L. E. & EISENBERG, E. (1980a) Dissociation of the actin-subfragment-one complex by adenylylimidodiphosphate, ADP and PPi.J. biol. Chem. 255, 543–8.Google Scholar
  13. GREENE, L. E. & EISENBERG, E. (1980b) The binding of heavy meromyosin to F-actin.J. biol. Chem. 255, 549–54.Google Scholar
  14. HOFMANN, W. & GOODY, R. S. (1978) The ternary complex formed between actin, myosin subfragment-1 and ATP (β,γ-NH).FEBS Lett. 89, 169–72.Google Scholar
  15. HUXLEY, A. F. (1957) Muscle structure and theories of contraction.Prog. biophys. molec. Biol. 7, 255–318.Google Scholar
  16. HUXLEY, H. E. (1971) The structural basis of muscular contraction.Proc. R. Soc. Lond. Ser. B 178, 131–49.Google Scholar
  17. KEGELES, G., RHODES, L. & BETHUNE, J. L. (1967) Sedimentation behaviour of chemically reacting systems.Proc. natn. Acad. Sci. U.S.A. 58, 45–51.Google Scholar
  18. KENYON, G. L. & BRUICE, T. W. C. (1977) Novel sulphydryl reagents. InMethods in Enzymology Vol. 47 (edited by HIRS, C. H. W. and TIMASHELF, S. N.), pp. 407–430. New York: Academic Press.Google Scholar
  19. KUHN, H. J. (1977) Reversible transformation of mechanical work into chemical free energy by stretch dependant binding of AMP-PNP in glycerinated fibrillar muscle fibres. InInsect Flight Muscle (edited by TREGEAR, R. T.), pp. 307–16. Amsterdam: North-Holland.Google Scholar
  20. LIN, T. (1978) Fluorimetric studies of actin labelled with dansyl aziridine.Archs Biochem. Biophys. 185, 285–99.Google Scholar
  21. LUSTY, G. J. & FASOLD, H. (1969) Characterization of sulphydryl groups of actin.Biochemistry 8, 2933–9.Google Scholar
  22. LYMN, R. W. (1976) Low angle X-ray diagrams from skeletal muscle: the effect of AMP.PNP, a non-hydrolysable analogue of ATP.J. molec. Biol. 99, 567–82.Google Scholar
  23. LYMN, R. W. & TAYLOR, E. W. (1971) Mechanism of adenosine triphosphate hydrolysis by actomyosin.Biochemistry 10, 4617–24.Google Scholar
  24. MARSTON, S. B., RODGER, C. D. & TREGEAR, R. T. (1976) Changes in muscle crossbridges when β-γ imido-ATP binds to myosin.J. molec. Biol. 104, 263–76.Google Scholar
  25. MARSTON, S. B., TREGEAR, R. T., RODGER, C. D. & CLARKE, M. L. (1979) Coupling between the enzymic site of myosin and the mechanical output of muscle.J. molec. Biol. 128, 111–26.Google Scholar
  26. MARSTON, S. B. & WEBER, A. (1975) The dissociation constant of the actin-heavy meromyosin subfragment-1 complex.Biochemistry 14, 3868–73.Google Scholar
  27. PERRY, S. V. (1955) Myosin adenosine triphosphate. InMethods in Enzymology Vol. 2 (edited by COLOWICK, S. P. and KAPLAN, N. O.), pp. 582–588. New York: Academic Press.Google Scholar
  28. PORTER, M. & WEBER, A. (1979) Non-cooperative response of actin-cysteine 373 in cooperatively behaving regulated actin filaments.FEBS Lett. 105, 259–62.Google Scholar
  29. REEDY, M. K., HOLMES, K. C. & TREGEAR, R. T. (1965) Induced changes in orientation of the crossbridges of glycerinated insect flight muscle.Nature 207, 1276–80.Google Scholar
  30. REES, M. K. & YOUNG, M. (1967) Studies on the isolation and molecular properties of homogenous globular actin.J. biol. Chem. 242, 4449–58.Google Scholar
  31. RODGER, C. D. & TREGEAR, R. T. (1974) Crossbridge angle when ADP is bound to myosin.J. molec. Biol. 86, 495–7.Google Scholar
  32. SCOUTEN, W. H., LUBCHER, R. & BAUGHMAN, W. (1974) N-dansyl aziridine: a new fluorescent modification for cysteine thiols.Biochim. biophys. acta 336, 421–6.Google Scholar
  33. SPUDICH, J. A. & WATT, S. (1971) The regulation of rabbit skeletal muscle contraction.J. biol. Chem. 246, 4866–71.Google Scholar
  34. STEIN, L. A., SCHWARZ, R. P., CHOCK, P. B. & EISENBERG, E. (1979) Mechanism of actomyosin ATPase.Biochemistry 18, 3895–909.Google Scholar
  35. STRAUB, F. B. (1942) Actin.Studies from the Institute of Medical Chemistry, University Szeged 2, 3–15.Google Scholar
  36. SWENSON, C. A. & RITCHIE, P. A. (1979) Enthalpy of nucleotides binding to myosin.Biochemistry 18, 3654–68.Google Scholar
  37. TANFORD, C. (1961)Physical Chemistry of Macromolecules. New York: Wiley.Google Scholar
  38. TREGEAR, R. T. & MARSTON, S. B. (1979) The crossbridge cycle.Ann. Rev. Physiol. 41, 723–36.Google Scholar
  39. TREGEAR, R. T., MILCH, J. R., GOODY, R. S., HOLMES, K. C. & RODGER, C. D. (1979) The use of some novel X-ray diffraction techniques to study the effects of nucleotides on crossbridges in insect flight muscle. InCrossbridge Mechanism in Muscle Contraction (edited by SUGI, H. and POLLACK, G. H.), pp. 407–423. Tokyo: University of Tokyo Press.Google Scholar
  40. WEEDS, A. G. & TAYLOR, R. S. (1975) Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin.Nature 252, 646–9.Google Scholar
  41. WEGNER, A. (1979) Equilibrium of actin-tropomyosin interaction.J. molec. Biol. 131, 839–53.Google Scholar
  42. WHITE, H. D. & TAYLOR, E. W. (1976) Energetics and mechanism of actomyosin adenosine triphosphatase.Biochemistry 15, 5818–26.Google Scholar

Copyright information

© Chapman and Hall Ltd 1980

Authors and Affiliations

  • S. B. Marston
    • 1
  1. 1.Biochemistry DepartmentICI Pharmaceuticals LtdMacclesfield

Personalised recommendations