Advertisement

Journal of Muscle Research and Cell Motility

, Volume 33, Issue 5, pp 305–312 | Cite as

Allosteric communication in Dictyostelium myosin II

  • Piyali Guhathakurta
  • Ewa Prochniewicz
  • Joseph M. Muretta
  • Margaret A. Titus
  • David D. Thomas
Original Paper

Abstract

Myosin’s affinities for nucleotides and actin are reciprocal. Actin-binding substantially reduces the affinity of ATP for myosin, but the effect of actin on myosin’s ADP affinity is quite variable among myosin isoforms, serving as the principal mechanism for tuning the actomyosin system to specific physiological purposes. To understand the structural basis of this variable relationship between actin and ADP binding, we studied several constructs of the catalytic domain of Dictyostelium myosin II, varying their length (from the N-terminal origin) and cysteine content. The constructs varied considerably in their actin-activated ATPase activity and in the effect of actin on ADP affinity. Actin had no significant effect on ADP affinity for a single-cysteine catalytic domain construct, a double-cysteine construct partially restored the actin-dependence of ADP binding, and restoration of all native Cys restored it further, but full restoration of function (similar to that of skeletal muscle myosin II) was obtained only by adding all native Cys and an artificial lever arm extension. Pyrene-actin fluorescence confirmed these effects on ADP binding to actomyosin. We conclude that myosin’s Cys content and lever arm both allosterically modulate the reciprocal affinities of myosin for ADP and actin, a key determinant of the biological functions of myosin isoforms.

Keywords

Actin Cysteine Lever arm ADP Fluorescence 

Notes

Acknowledgments

Fluorescence measurements were performed in the Biophysical Spectroscopy Facility, University of Minnesota. We thank Sarah Blakeley, Christina Yi, and Evan Smith for technical assistance and to Octavian Cornea for assistance with manuscript preparation. This work was supported by NIH grants to DDT (AR32961 and AG26160), and PG was supported by the Minnesota Muscle Training Grant, NIH T32 AR007612.

References

  1. Agafonov RV, Nesmelov YE, Titus MA, Thomas DD (2008) Muscle and nonmuscle myosins probed by a spin label at equivalent sites in the force-generating domain. Proc Natl Acad Sci USA 105(36):13397–13402PubMedCrossRefGoogle Scholar
  2. Agafonov RV, Negrashov IV, Tkachev YV, Blakely SE, Titus MA, Thomas DD, Nesmelov YE (2009) Structural dynamics of the myosin relay helix by time-resolved EPR and FRET. Proc Natl Acad Sci USA 106(51):21625–21630PubMedCrossRefGoogle Scholar
  3. Batra R, Geeves MA, Manstein DJ (1999) Kinetic analysis of Dictyostelium discoideum myosin motor domains with glycine-to-alanine mutations in the reactive thiol region. Biochemistry 38(19):6126–6134PubMedCrossRefGoogle Scholar
  4. Bauer CB, Kuhlman PA, Bagshaw CR, Rayment I (1997) X-ray crystal structure and solution fluorescence characterization of Mg.2′(3′)-O-(N-methylanthraniloyl) nucleotides bound to the Dictyostelium discoideum myosin motor domain. J Mol Biol 274(3):394–407PubMedCrossRefGoogle Scholar
  5. Berg JS, Powell BC, Cheney RE (2001) A millennial myosin census. Mol Biol Cell 12(4):780–794PubMedGoogle Scholar
  6. Bloemink MJ, Geeves MA (2011) Shaking the myosin family tree: biochemical kinetics defines four types of myosin motor. Semin Cell Dev Biol 22(9):961–967PubMedCrossRefGoogle Scholar
  7. Cremo CR, Geeves MA (1998) Interaction of actin and ADP with the head domain of smooth muscle myosin: implications for strain-dependent ADP release in smooth muscle. Biochemistry 37(7):1969–1978PubMedCrossRefGoogle Scholar
  8. Cremo CR, Neuron JM, Yount RG (1990) Interaction of myosin subfragment 1 with fluorescent ribose-modified nucleotides. A comparison of vanadate trapping and SH1-SH2 cross-linking. Biochemistry 29(13):3309–3319PubMedCrossRefGoogle Scholar
  9. Criddle AH, Geeves MA, Jeffries T (1985) The use of actin labelled with N-(1-pyrenyl)iodoacetamide to study the interaction of actin with myosin subfragments and troponin/tropomyosin. Biochem J 232(2):343–349PubMedGoogle Scholar
  10. De La Cruz EM, Ostap EM (2009) Kinetic and equilibrium analysis of the myosin ATPase. Methods Enzymol 455:157–192CrossRefGoogle Scholar
  11. De La Cruz EM, Wells AL, Rosenfeld SS, Ostap EM, Sweeney HL (1999) The kinetic mechanism of myosin V. Proc Natl Acad Sci USA 96(24):13726–13731CrossRefGoogle Scholar
  12. De La Cruz EM, Sweeney HL, Ostap EM (2000) ADP inhibition of myosin V ATPase activity. Biophys J 79(3):1524–1529CrossRefGoogle Scholar
  13. De La Cruz EM, Ostap EM, Sweeney HL (2001) Kinetic mechanism and regulation of myosin VI. J Biol Chem 276(34):32373–32381CrossRefGoogle Scholar
  14. Foth BJ, Goedecke MC, Soldati D (2006) New insights into myosin evolution and classification. Proc Natl Acad Sci USA 103(10):3681–3686PubMedCrossRefGoogle Scholar
  15. Franks-Skiba K, Cooke R (1995) The conformation of the active site of myosin probed using mant-nucleotides. Biophys J 68(4 Suppl):142S–147S (discussion 147S–149S)PubMedGoogle Scholar
  16. Geeves MA (1989) Dynamic interaction between actin and myosin subfragment 1 in the presence of ADP. Biochemistry 28(14):5864–5871PubMedCrossRefGoogle Scholar
  17. Geeves MA, Jeffries TE, Millar NC (1986) ATP-induced dissociation of rabbit skeletal actomyosin subfragment 1. Characterization of an isomerization of the ternary acto-S1-ATP complex. Biochemistry 25(26):8454–8458PubMedCrossRefGoogle Scholar
  18. Greene LE, Eisenberg E (1980) Dissociation of the actin.subfragment 1 complex by adenyl-5′-yl imidodiphosphate, ADP, and PPi. J Biol Chem 255(2):543–548PubMedGoogle Scholar
  19. Himmel DM, Mui S, O’Neall-Hennessey E, Szent-Gyorgyi AG, Cohen C (2009) The on-off switch in regulated myosins: different triggers but related mechanisms. J Mol Biol 394(3):496–505PubMedCrossRefGoogle Scholar
  20. Hiratsuka T (1984) Distinct structures of ATP and GTP complexes in the myosin ATPase. J Biochem 96(1):155–162PubMedGoogle Scholar
  21. Holmes KC, Angert I, Kull FJ, Jahn W, Schroder RR (2003) Electron cryo-microscopy shows how strong binding of myosin to actin releases nucleotide. Nature 425(6956):423–427PubMedCrossRefGoogle Scholar
  22. Houdusse A, Cohen C (1996) Structure of the regulatory domain of scallop myosin at 2 A resolution: implications for regulation. Structure 4(1):21–32PubMedCrossRefGoogle Scholar
  23. Kast D, Espinoza-Fonseca LM, Yi C, Thomas DD (2010) Phosphorylation-induced structural changes in smooth muscle myosin regulatory light chain. Proc Natl Acad Sci USA 107(18):8207–8212PubMedCrossRefGoogle Scholar
  24. Kintses B, Gyimesi M, Pearson DS, Geeves MA, Zeng W, Bagshaw CR, Malnasi-Csizmadia A (2007) Reversible movement of switch 1 loop of myosin determines actin interaction. EMBO J 26(1):265–274PubMedCrossRefGoogle Scholar
  25. Korman VL, Anderson SE, Prochniewicz E, Titus MA, Thomas DD (2006) Structural dynamics of the actin-myosin interface by site-directed spectroscopy. J Mol Biol 356(5):1107–1117PubMedCrossRefGoogle Scholar
  26. Kouyama T, Mihashi K (1981) Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labelled F-actin. Local structural change of actin protomer both on polymerization and on binding of heavy meromyosin. Eur J Biochem 114(1):33–38PubMedCrossRefGoogle Scholar
  27. Kurzawa SE, Manstein DJ, Geeves MA (1997) Dictyostelium discoideum myosin II: characterization of functional myosin motor fragments. Biochemistry 36(2):317–323PubMedCrossRefGoogle Scholar
  28. Lanzetta PA, Alvarez LJ, Reinach PS, Candia OA (1979) An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem 100(1):95–97PubMedCrossRefGoogle Scholar
  29. Lowey S, Waller GS, Trybus KM (1993) Skeletal muscle myosin light chains are essential for physiological speeds of shortening. Nature 365(6445):454–456PubMedCrossRefGoogle Scholar
  30. Manstein DJ, Hunt DM (1995) Overexpression of myosin motor domains in Dictyostelium: screening of transformants and purification of affinity tagged protein. J Muscle Res Cell Motil 16:325–332PubMedCrossRefGoogle Scholar
  31. Mello R, Thomas DD (2012) Three distinct actin-attached structural states of myosin in muscle fibers. Biophys J 102:1088-1096Google Scholar
  32. Nyitrai M, Geeves MA (2004) Adenosine diphosphate and strain sensitivity in myosin motors. Philos Trans R Soc Lond B Biol Sci 359(1452):1867–1877PubMedCrossRefGoogle Scholar
  33. O’Connell CB, Tyska MJ, Mooseker MS (2007) Myosin at work: motor adaptations for a variety of cellular functions. Biochim Biophys Acta 1773(5):615–630PubMedCrossRefGoogle Scholar
  34. Prochniewicz E, Zhang Q, Howard EC, Thomas DD (1996) Microsecond rotational dynamics of actin: spectroscopic detection and theoretical simulation. J Mol Biol 255(3):446–457PubMedCrossRefGoogle Scholar
  35. Ritchie MD, Geeves MA, Woodward SK, Manstein DJ (1993) Kinetic characterization of a cytoplasmic myosin motor domain expressed in Dictyostelium discoideum. Proc Natl Acad Sci USA 90(18):8619–8623PubMedCrossRefGoogle Scholar
  36. Siemankowski RF, Wiseman MO, White HD (1985) ADP dissociation from actomyosin subfragment 1 is sufficiently slow to limit the unloaded shortening velocity in vertebrate muscle. Proc Natl Acad Sci USA 82(3):658–662PubMedCrossRefGoogle Scholar
  37. Sweeney HL, Houdusse A (2004) The motor mechanism of myosin V: insights for muscle contraction. Philos Trans R Soc Lond B Biol Sci 359(1452):1829–1841PubMedCrossRefGoogle Scholar
  38. Thomas DD, Kast D, Korman VL (2009) Site-directed spectroscopic probes of actomyosin structural dynamics. Annu Rev Biophys 38:347–369PubMedCrossRefGoogle Scholar
  39. Wagner PD, Slater CS, Pope B, Weeds AG (1979) Studies on the actin activation of myosin subfragment-1 isoezymes and the role of myosin light chains. Eur J Biochem 99(2):385–394PubMedCrossRefGoogle Scholar
  40. Wells JA, Yount RG (1979) Active site trapping of nucleotides by crosslinking two sulfhydryls in myosin subfragment 1. Proc Natl Acad Sci USA 76(10):4966–4970PubMedCrossRefGoogle Scholar
  41. Whittaker M, Wilson-Kubalek EM, Smith JE, Faust L, Milligan RA, Sweeney HL (1995) A 35-A movement of smooth muscle myosin on ADP release. Nature 378(6558):748–751PubMedCrossRefGoogle Scholar
  42. Woodward SK, Eccleston JF, Geeves MA (1991) Kinetics of the interaction of 2′(3′)-O-(N-methylanthraniloyl)-ATP with myosin subfragment 1 and actomyosin subfragment 1: characterization of two acto-S1-ADP complexes. Biochemistry 30(2):422–430PubMedCrossRefGoogle Scholar
  43. Woodward SK, Geeves MA, Manstein DJ (1995) Kinetic characterization of the catalytic domain of Dictyostelium discoideum myosin. Biochemistry 34(49):16056–16064PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Piyali Guhathakurta
    • 1
  • Ewa Prochniewicz
    • 1
  • Joseph M. Muretta
    • 1
  • Margaret A. Titus
    • 2
  • David D. Thomas
    • 1
  1. 1.Department of Biochemistry, Molecular Biology, and BiophysicsUniversity of MinnesotaMinneapolisUSA
  2. 2.Department of Genetics, Cell Biology, and DevelopmentUniversity of MinnesotaMinneapolisUSA

Personalised recommendations