Studies on the ATP-binding Site of Actin Using Site-directed Mutagenesis

  • Herwig Schüler
  • Elena Korenbaum
  • Uno Lindberg
  • Roger Karlsson
Part of the NATO ASI Series book series (volume 102)


Hydrolysis of the actin-bound AT? is linked to actin filament turnover. However, neither the role nor the mechanism of the actin ATPase are well established. A novel role for the actin ATPase in muscle contraction has been proposed, making actin the principal force generator (1). The crystal structures of α- (2,3) and β-actin (4) suggest involvement of serine 14 (S14) and aspartic acid 157 (D157) in ATP hydrolysis (2,5). Myslik (5) suggested that the serine hydroxyl, polarized by the aspartic carboxyl, might be transiently phosphorylated under the hydrolysis reaction. According to his analysis (5) of the β-actin structure (4), the geometry of the ATP-site would favor an in-line attack on the ATPγP by the serine hydroxyl. This model was also inspired by the geometric homology of actin to the ATPase domain of Hsc70, which places alcoholic and acidic side chains at similar positions (6).


ATPase Activity Actin Filament Motility Assay Myosin Binding Heat Shock Cognate Protein 
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  1. 1.
    Schutt, C.E. & Lindberg, U. (1992), Actin as the generator of force during muscle contraction. PNAS 89, 319–323PubMedCrossRefGoogle Scholar
  2. 2.
    Kabsch, W. et al(1990), Atomic structure of the actin:DNase I complex. Nature 347, 37–44PubMedCrossRefGoogle Scholar
  3. 3.
    McLaughlin, P.J. et al(1993), Structure of gelsolin segment 1-actin complex and the mechanism of filament severing. Nature 364, 685–692PubMedCrossRefGoogle Scholar
  4. 4.
    Schutt, C.E. et al(1993), The structure of crystalline profilin-β-actin. Nature 365, 810–816PubMedCrossRefGoogle Scholar
  5. 5.
    Myslik, J.C. (1992), The structure of actin in the crystalline profiling:actin complex. Thesis, Princeton UniversityGoogle Scholar
  6. 6.
    Flaherty, K.M. et al(1991), Similarity of the three-dimensional structures of actin and the ATPase fragment of a 70-kDa heat shock cognate protein. PNAS 88, 5041–5045PubMedCrossRefGoogle Scholar
  7. 7.
    Aspenström, P. et al(1993), Mutations in β-actin: influence on polymer formation and on interactions with myosin and profilin. FEBS letters 329, 163–170PubMedCrossRefGoogle Scholar
  8. 8.
    Chen, X. & Rubenstein, P.A. (1995), A mutation in an ATP-binding loop of Saccharomyces cerevisiaeactin (Sl4A) causes a temperature-sensitive phenotype in vivoand in vitroJ Biol Chem 270, 11406–11414PubMedCrossRefGoogle Scholar
  9. 9.
    Karlsson, R. (1988), Expression of chicken beta-actin in Saccharomyces cerevisiaeGene 68, 249–257PubMedCrossRefGoogle Scholar
  10. 10.
    Kouyama, T. & Mihashi, K. (1981), Fluorimetry study of N-(1-pyrenyl)iodoacetamide labelled F-actin. Eur J Biochem 114, 33–38PubMedCrossRefGoogle Scholar
  11. 11.
    Blikstad, I. et al(1978), Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell 15, 935–943PubMedCrossRefGoogle Scholar
  12. 12.
    Seals, J.R. et al(1978), A sensitive and precise isotopic assay of ATPase activity. Anal Biochem 90, 785–795PubMedCrossRefGoogle Scholar
  13. 13.
    Spudich, J.A. (1974), Biochemical and structural studies of actomyosin-like proteins from non-muscle cells. J Biol Chem 249, 6013–6020PubMedGoogle Scholar
  14. 14.
    Kron, S.J. & Spudich, J.A. (1986), Fluorescent actin filaments move on myosin fixed to a glass surface. PNAS 83, 6272–6276.PubMedCrossRefGoogle Scholar
  15. 15.
    Kishino, A. & Yanagida, T. (1988), Force measurement by micromanipulation of a single actin filament by glass needles. Nature 334, 74–76.PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1997

Authors and Affiliations

  • Herwig Schüler
    • 1
  • Elena Korenbaum
    • 1
  • Uno Lindberg
    • 1
  • Roger Karlsson
    • 1
  1. 1.Department of Cell Biology, The Wenner-Gren InstituteStockholm UniversityStockholmSweden

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