Advertisement

Actin pp 59-70 | Cite as

C-Terminus on Acitn: Spectroscopic and Immunochemical Examination of its Role in Actomyosin Interactions

  • Anh M. Duong
  • Emil Reisler
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 358)

Abstract

The understanding of force generation during cyclic interactions of myosin with actin requires a detailed description of actomyosin interface and its evolution during ATP hydrolysis. Until now, driven by the availability of mutants1–4 and atomic resolution structure of actin5, the mapping of actomyosin interface has focused on the determination of myosin binding sites on actin. Two proximal areas on actin, in the N- and C-terminal regions of this protein, have been implicated by structural considerations5,6 and biochemical7,8, immunochemical, NMR, and mutagenic studies1–3,15 in the binding of myosin heads (S-1). Immunochemical10and mutagenic approaches16 suggested also the involvement of residues 91–103 on actin in actomyosin interactions. While the previous studies produced a general agreement on the important contribution of N-terminal acidic residues 1–4 on actin to the activation of the myosin ATPase activity and the motility of actin filaments, the actual role of actin’s C-terminal residues in actomyosin interactions has not been assessed. An interesting approach to this task, which was employed also in the work on LC-2 myosin light chains17, was taken by Labbe et al.10 These authors modified the penultimate cysteine residue on actin, Cys-374, with N-iodoacetyl-N’-(5-sulpho-1-naphtyl)ethylenediamine (1,5- IAEDANS) and showed in solid phase immunochemical assays (ELISA) that S-1 and antidansyl antibodies competed with each other for the binding to the modified cysteine.

Keywords

ATPase Activity Myosin Head Myosin ATPase Activity Actin Concentration Myosin Subfragment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    R.K. Cook, W.T. Blake, and P.A. Rubenstein, Removal of the amino-terminal acidic residues of yeast actin, J. Biol. Chem., 267: 9430 (1992).PubMedGoogle Scholar
  2. 2.
    P. Aspenstrom, and R. Karlsson, Interference with myosin subfragment-1 binding by site-directed mutagenesis of actin, Eur. J. Biochem., 200: 35 (1992).Google Scholar
  3. 3.
    K. Sutoh, M. Ando, K. Sutoh, and Y.Y. Toyoshima, Site-directed mutations in Dictyostelium actin: disruption of a negative charge cluster at the N-terminus, Proc. Natl. Acad. Sci. USA, 88: 7711 (1991).PubMedCrossRefGoogle Scholar
  4. 4.
    K.F. Wertman, D.G. Drubin, and D. Botstein, Systematic mutational analysis of the yeast ACT 1 gene, Genetics, 132: 337 (1992).PubMedGoogle Scholar
  5. 5.
    W. Kabsch, H.G. Mannherz, D. Suck, E. Pai, and K.C. Holmes, Atomic structure of the actin-DNase I complex, Nature, 347: 37 (1990).PubMedCrossRefGoogle Scholar
  6. 6.
    W. Kabsch, and J. Vandekerchove, Structure and function of actin, Annu. Rev. Biophys. Biomol. Struct., 21: 49 (1992).PubMedCrossRefGoogle Scholar
  7. 7.
    R. Bertrand, P. Chaussepied, E. Audemard, and R. Kassab, Functional characterization of skeletal F-actin labeled on the NH2-terminal segment of residues 1-28, Eur. J. Biochem., 181: 747 (1989).CrossRefGoogle Scholar
  8. 8.
    J.E. Van Eyk, and R. Hodges, A synthetic peptide of the N-terminus of actin interacts with myosin, Biochemistry, 30: 11676 (1991).CrossRefGoogle Scholar
  9. 9.
    C. Mejean, M. Boyer, J.P. Labbe, J. Derancourt, Y. Benyamin, and C. Roustan, Anti-actin antibodies: an immunological approach to the myosin-actin and tropomyosin-actin interfaces, Biochem. J., 244: 571 (1987).Google Scholar
  10. 10.
    J.P. Labbe, C. Mejean, Y. Benyamin, and C. Roustan, Characterization of an actin-myosin head interface in the 40-113 region of actin using specific antibodies as probes Biochem. J., 271: 407 (1990).Google Scholar
  11. 11.
    L. Miller, M. Kalnoski, Z. Yunossi, J.C. Bulinski, and E. Reisler, Antibodies directed against N-terminal residues on actin do not block acto-myosin binding, Biochemistry, 26: 6064 (1987).CrossRefGoogle Scholar
  12. 12.
    G. DasGupta, and E. Reisler, Antibody against the amino terminus of α-actin inhibits actomyosin interactions in the presence of ATP, J. Mol. Biol., 207: 833 (1989).CrossRefGoogle Scholar
  13. 13.
    G. DasGupta, and E. Reisler, Acto-myosin interactions in the presence of ATP and the N-terminal segment of actin, Biochemistry, 31: 1836 (1992).CrossRefGoogle Scholar
  14. 14.
    A.J.G. Moir, and B. A. Levine, Protein cognitive sites on the surface of actin. A proton NMR study, J. Inorganic Chem., 27: 271 (1986).Google Scholar
  15. 15.
    R.K. Cook, D. Root, C. Miller, E. Reisler, and P.A. Rubenstein, Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus, J. Biol. Chem., 268: 2410 (1993).Google Scholar
  16. 16.
    M. Johara, Y.Y. Toyoshima, A. Ishijima, H. Kojima, T. Yanagida, and K. Sutoh, Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion, Proc. Natl. Acad. Sci. USA, 90: 2127 (1993).CrossRefGoogle Scholar
  17. 17.
    T. Katoh, and S. Lowey, Mapping myosin light chains by immunoelectron microscopy. Use of anti-fluorescyl antibodies as structural probes, J. Cell Biol., 109: 1549 (1989).CrossRefGoogle Scholar
  18. 18.
    B. Malm, L.E. Nystrom, and U. Lindberg, The effect of proteolysis on the stability of the profilactin complex, FEBS Lett., 113: 241 (1980).CrossRefGoogle Scholar
  19. 19.
    P. Graceffa, and A. Jancso, Disulfide cross-linking of caldesmon to actin, J. Biol. Chem., 266: 20305 (1991).Google Scholar
  20. 20.
    Y. Doi, M. Banba, and A. Vertut-Doi, Cysteine 374 of actin resides at the gelsolin contact site in the EGTA resistant actin-gelsolin complex, Biochemistry, 30: 5769 (1991).CrossRefGoogle Scholar
  21. 21.
    C. Combeau, D. Didry, and M.-F. Carlier, Interaction between G-actin and myosin subfragment-1 probed by covalent cross-linking, J. Biol. Chem., 267: 14038 (1992).Google Scholar
  22. 22.
    R.H. Crosbie, J.M. Chalovich, and E. Reisler, Interaction of caldesmon and myosin subfragment 1 with the C-terminus of actin, Biochem. Biophys. Res. Commun., 184: 239 (1992).CrossRefGoogle Scholar
  23. 23.
    R. Makuch, J. Kotakowski, and R. Dabrowska, The importance of C-terminal amino acid residues of actin to the inhibition of actomyosin ATPase activity by caldesmon and troponin I, FEBS Lett., 297: 237 (1992).CrossRefGoogle Scholar
  24. 24.
    S.I. O’Donoghue, M. Miki, and C.G. DosRemidios, Removing the two C-terminal residues of actin affects the filament structure, Arch. Biochem. Biophys. 293: 110 (1992).CrossRefGoogle Scholar
  25. 25.
    A.G. Weeds, and B. Pope, Studies on the chymotryptic digestion of myosin. Effects of divalent cations on proteolytic susceptibility, J. Mol. Biol., 111: 129 (1977).CrossRefGoogle Scholar
  26. 26.
    J.A. Spudich, and S. Watt, Regulation of skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin, J. Biol. Chem., 246: 4866 (1971).Google Scholar
  27. 27.
    D.E. Lopatin, and E.W. Voss, Jr., Fluorescein. Hapten and antibody active-site probe, Biochemistry, 10: 208 (1971).CrossRefGoogle Scholar
  28. 28.
    R. Takahashi, Fluorescence energy transfer between subfragment-1 and actin points in the rigor complex of acto subfragment-1, Biochemistry, 18: 5164 (1979).CrossRefGoogle Scholar
  29. 29.
    T.-I. Lin, Fluorimetric studies of actin-labeled with dansyl aziridine, Arch. Biochem. Biophys., 185: 285 (1978).CrossRefGoogle Scholar
  30. 30.
    T. Tao, and J. Cho, Fluorescence lifetime quenching studies on the accessibilities of actin sulfhydryl sites, Biochemistry, 18: 2759 (1979).CrossRefGoogle Scholar
  31. 31.
    L. Miller, M. Phillips, and E. Reisler, Polymerization of G-actin by myosin subfragment 1, J. Biol. Chem., 263: 1996 (1988).Google Scholar
  32. 32.
    U.K. Laemmli, Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature (London) 227: 680 (1970).CrossRefGoogle Scholar
  33. 33.
    J. Reidler, V.T. Oi, W. Carlsen, T.M. Vuong, I. Pecht, L.A. Herzenberg, and L. Stryer, Rotational dynamics of monoclonal anti-dansyl immunoglobulins, J. Mol. Biol., 158: 739 (1982).CrossRefGoogle Scholar
  34. 34.
    G. DasGupta, and E. Reisler, Nucleotide-induced changes in the interaction of myosin subfragment 1 with actin: Detection by antibodies against the N-terminal segment of actin, Biochemistry, 30: 9961 (1991).CrossRefGoogle Scholar
  35. 35.
    E. Reisler, Actin molecular structure and function, Curr. Opin. Cell Biol., 5: 41 (1993).CrossRefGoogle Scholar
  36. 36.
    T. Kouyama, and K. Mihashi, Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labeled F-actin, Eur. J. Biochem. 114: 33 (1981).Google Scholar
  37. 37.
    M. Mossakowska, J. Moraczewska, S. Khaitlina, H. Strzelecka-Golaszewska, Proteolytic removal of three C-terminal residues of actin alters the monomer-monomer interactions, Biochem. J. 289: 897 (1992).Google Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Anh M. Duong
    • 1
  • Emil Reisler
    • 1
  1. 1.Department of Chemistry and Biochemistry and the Molecular Biology InstituteUniversity of CaliforniaLos AngelesUSA

Personalised recommendations