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
Chapter
Part of the Topics in Molecular and Structural Biology book series (TMSB)

Abstract

The structure of the G-actin monomer complexed with DNase I has been solved by X-ray crystallography to a resolution of 0.6 nm (Figure 3.1; Suck et al., 1981). More recently the resolution has been increased to 0.45 nm and the actin molecule has been unambiguously identified (Kabsch et al., 1985). Work at present in progress in our laboratory has extended the resolution to 0.3 nm. This resolution has revealed the course of the chain in the actin monomer. Below, we describe the structure of F-actin arrived at by the following search procedure: the structure obtained from crystallography at 0.6 nm is placed in all possible orientations in the F-actin helix; the radius is adjusted to give the observed radius of gyration (2.5 nm: Hartt and Mendelson, 1980); the fibre diffraction pattern is computed from the resulting structure and compared with the X-ray diffraction data from oriented gels of F-actin (Popp et al., 1987) measured to 0.8 nm resolution. The intensity R-factor was used as an index of fit:
$$R = \frac{{\sum w{{\left( {{I_{calc}} - {I_{obs}}} \right)}^2}}}{{\sum w{I_{obs}}^2}}$$

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References

  1. Aebi, U., Millonig, R., Salvo, H. and Engel, A. (1986). The three-dimensional structure of the actin filament revisited. Ann. N. Y. Acad. Sci., 483, 100–119PubMedCrossRefGoogle Scholar
  2. Amos, L. A., Huxley, H. E., Holmes, K. C., Goody, R. S. and Taylor, K. A. (1982). Structural evidence that myosin heads may interact with two sites on F-actin. Nature, 299, 467–469PubMedCrossRefGoogle Scholar
  3. Egelman, E. H. and DeRosier, D. J. (1983a). A model for F-actin derived from image analysis of isolated filaments. J. Mol. Biol., 166, 623–629CrossRefGoogle Scholar
  4. Egelman, E. H. and DeRosier, D. J. (1983b). Structural studies of F-actin. In dos Remedios, C. and Barden, J. (Eds.), Actin: Its Structure and Function in Muscle and Non-muscle cells. Academic Press, London, pp. 17–24Google Scholar
  5. Hartt, J. and Mendelson, R. (1980). X-ray scattering of F-actin and myosin subfragment-1 complex. Fed. Proc, 39, 1728Google Scholar
  6. Holmes, K. C., Popp, D., Gebhard, W., Kabsch, W., Jahn, W. and Bryan, R. (1989). The structure of F-actin as revealed by analysis of the x-ray fibre diffraction pattern (in preparation)Google Scholar
  7. Kabsch, W., Mannherz, H. G. and Suck, D. (1985). Three-dimensional structure of the complex of actin and DNase I at 4.5 Å resolution. EMBO Jl, 4, 2113–2118Google Scholar
  8. Milligan, R. A. and Flicker, P. F. (1987). Structural relationships of actin, myosin, and tropomyosin revealed by cryo-electron microscopy. J. Cell Biol., 105, 29–39PubMedCrossRefGoogle Scholar
  9. Moore, P. B., Huxley, H. E. and DeRosier, D. J. (1970). Three-dimensional reconstruction of F-actin, thin filaments, and decorated thin filaments. J. Mol. Biol., 50, 279–295PubMedCrossRefGoogle Scholar
  10. O’Brien, E. J., Couch, J., Johnson, G. R. P. and Morris, E. P. (1983). Structure of actin and the thin filament. In dos Remedios, C. and Barden, J. (Eds.), Actin: Its Structure and Function on Muscle and Non-muscle Cells. Academic Press, London, pp. 3–16Google Scholar
  11. Popp, D. (1986). Röntgenstrukturuntersuchungen an orientierten F-actinfasern. Diploma Thesis, University of HeidelbergGoogle Scholar
  12. Popp, D. and Holmes, K. C. (1989). X-ray diffraction studies of vertebrate smooth muscle thin filaments (in preparation)Google Scholar
  13. Popp, D., Lednev, V. V. and Jahn, W. (1987). Methods of preparing well-orientated sols of F-actin containing filaments suitable for x-ray diffraction. J. Mol. Biol., 197, 679–684PubMedCrossRefGoogle Scholar
  14. Schutt, C., Strauss, N. and Lindberg, U. (1985). Crystallographic studies of the profilin-actin complex. J. Muscle Res. Cell Motil., 6, 668Google Scholar
  15. Spudich, J. A., Huxley, H. E. and Finch, J. (1972). Regulation of skeletal muscle contraction. II. Structural studies of the interactions of the troponin-tropomyosin complex with actin. J. Mol. Biol., 72, 619–632PubMedCrossRefGoogle Scholar
  16. Suck, D., Kabsch, W. and Mannherz, H. G. (1981). Three-dimensional structure of the complex of skeletal muscle actin and bovine pancreatic DNase I at 6-A resolution. Proc. Natl Acad. Sci. USA, 78, 4319–4323PubMedPubMedCentralCrossRefGoogle Scholar
  17. Sutoh, Kazuo (1982). Identification of myosin binding sites on the actin sequence. Biochemistry, 21, 3654–3661PubMedCrossRefGoogle Scholar
  18. Taylor, D. L., Reidler, J., Spudich, J. A. and Stryer, L. (1981). Detection of actin assembly by fluorescence energy transfer. J. Cell Biol., 89, 362–367PubMedCrossRefGoogle Scholar
  19. Trinick, J., Cooper, J., Seymour, J. and Egelman, E. H. (1986). Cryo-electron microscopy and three-dimensional reconstruction of actin filaments. J. Microsc., 141, 349–360PubMedCrossRefGoogle Scholar
  20. Wakabayashi, T., Huxley, H. E., Amos, L. and Klug, A. (1975). Three-dimensional image reconstruction of actin-tropomyosin complex and actin-tropomyosin-troponin I complex. J. Mol. Biol., 93, 477–497PubMedCrossRefGoogle Scholar

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© The editor and contributors 1990

Authors and Affiliations

  • K. C. Holmes
  • D. Popp
  • W. Gebhard
  • W. Kabsch

There are no affiliations available

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