The Structure of F-Actin Calculated from X-Ray Fibre Diagrams and the 0.6 nm Crystal Structure

  • K. C. Holmes
  • D. Popp
  • W. Gebhard
  • W. Kabsch
Conference paper
Part of the Springer Series in Biophysics book series (BIOPHYSICS, volume 3)

Abstract

The structure of the g-actin monomer complexed with DNaseI has been solved by x-ray crystallography to 0.45nm resolution, Fig.l (Kabsch, Mannherz, & Suck, 1985). In the following we describe the structure of f-actin arrived at by a search procedure: the structure obtained from crystallography at 0.6nm is placed in all possible orientations in the F-actin helix; the fibre diffraction pattern is computed from the resulting structure and compared with the x-ray diffraction data from orientated gels of F-actin (Popp, Lednev, & Jahn, 1986) measured to 0.8 nm resolution. This process yielded five possible solutions at low resolution (2.0nm) only one of which successfully refined to high resolution (0.8nm). A full account of this study is in preparation (Holmes, et al. 1989). The best of the five possible solutions resulting from the low resolution search is shown in Fig. 2. The (intensity) R-factor is 0.12. To refine at high resolution we adopted an iterative least squares procedure. However, no solution would refine satisfactorily. The resulting R-factors ranged from 0.37–0.45.

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References

  1. Aebi, U., Millonig, R., Salvo, H., & Engel, A. (1986) The three- dimensional structure of the actin filament revisited. Annal. New York Acad. Sci 483, 100–119.CrossRefGoogle Scholar
  2. Amos, L. A., Huxley, H. E., Holmes, K. C., Goody, R. S., & Taylor, K. A. (1982) Structural evidence that myosin heads may interact with two sites on F-actin. Nature (Lond.) 5882, 467–469.Google Scholar
  3. Egelman, E. H. & DeRosier, D. J. (1983). Structural studies of F- actin. in Actin: Its structure and function on muscle and non- muscle cells (ed. dos Remedios & Barden) pp 17–24. Academic Press London.Google Scholar
  4. Holmes, K. C., Popp, D., Gebhard, W., Kabsch, W., Jahn, W., & Bryan,R. (1989) The structure of f-actin as revealed by analysis of the x- ray fibre diffraction pattern, (in preparation)Google Scholar
  5. Kabsch, W., Mannherz, H. G. & Suck, D. (1985) Three dimensional structure of the complex of actin and DNase I at 4.5 A resolution. EMBO Journal 4, 2113–2118.PubMedGoogle Scholar
  6. Milligan, R. A. & Flicker, P.F. (1987) Structural relationships of actin, myosin, and tropomyosin revealed by cryo-electron microscopy. J. Cell Biology 105, 29–39CrossRefGoogle Scholar
  7. O’Brien, E. J., Couch, J., Johnson, G. R. P., & Morris, E. P. (1983) Structure of actin and the thin filament, in Actin: Its structure and function on muscle and non-muscle cells (ed. dos Remedios & Barden ) pp 3–16. Academic Press, London.Google Scholar
  8. Popp, D., Lednev, V. V., & Jahn, W. (1987) Methods of preparing well-orientated sols of f-actin containing filaments suitable for x-ray diffraction. J. molec. Biol. 197, 679–684PubMedCrossRefGoogle Scholar
  9. Sutoh, K. (1982) Identification of myosin binding sites on the actin sequence. Biochemistry 21, 3654–3661PubMedCrossRefGoogle Scholar
  10. Taylor, D. L., Reidler, J., Spudich, J. A., & Stryer, L. (1981) Detection of actin assembly by fluorescence energy transfer. J. Cell Biology 89, 362–367.CrossRefGoogle Scholar
  11. Trinick, J., Cooper, J., Seymour, J., & Egelman, E.H. (1986) Cryoelectron microscopy and three-dimensional reconstruction of actin filaments. J. of Microscopy 141, 349–360.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1989

Authors and Affiliations

  • K. C. Holmes
    • 1
  • D. Popp
    • 1
  • W. Gebhard
    • 1
  • W. Kabsch
    • 1
  1. 1.Max Planck Institut für medizinische ForschungHeidelbergW. Germany

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