Journal of Muscle Research & Cell Motility

, Volume 25, Issue 4–5, pp 337–341 | Cite as

Specificity of blebbistatin, an inhibitor of myosin II

  • John Limouze
  • Aaron F. Straight
  • Timothy Mitchison
  • James R. Sellers


Blebbistatin is a small molecule inhibitor discovered in a screen for inhibitors of nonmuscle myosin IIA. We have examined the specificity and potency of the drug by assaying its effects on the actin-activated MgATPase assay of diverse members of the myosin superfamily. Blebbistatin potently inhibits several striated muscle myosins as well as vertebrate nonmuscle myosin IIA and IIB with IC50 values ranging from 0.5 to 5 μM. Interestingly, smooth muscle which is highly homologous to vertebrate nonmuscle myosin is only poorly inhibited (IC50=80 μM). The drug potently inhibits Dictyostelium myosin II, but poorly inhibits Acanthamoeba myosin II. Blebbistatin did not inhibit representative myosin superfamily members from classes I, V, and X.

myosin blebbistatin inhibitors 


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  1. Adelstein RS and Klee CB (1981) Purification and characterization of smooth muscle myosin light chain kinase. J Biol Chem 256: 7501–7509.PubMedGoogle Scholar
  2. Avraham KB (2003) Molecular motors in sensory defects. In: Schliwa M. (ed.) Molecular Motors. (pp. 511–537) Wiley-VCH, Wenheim.Google Scholar
  3. Berg JS, Powell BC and Cheney RE (2001) A millennial myosin census. Mol Biol Cell 12: 780–794.PubMedGoogle Scholar
  4. Brzeska H and Korn ED (1996) Regulation of class I and class II myosins by heavy chain phosphorylation. J Biol Chem 271: 16983–16986.PubMedCrossRefGoogle Scholar
  5. Cheung A, Dantzig JA, Hollingworth S, Baylor SM, Goldman YE, Mitchison TJ and Straight AF (2002) A small-molecule inhibitor of skeletal muscle myosin II. Nature Cell Biology 4: 83–88.PubMedCrossRefGoogle Scholar
  6. Cuda G, Pate E, Cooke R and Sellers JR (1997) In vitro actin filament sliding velocities produced by mixtures of different types of myosin. Biophys J 72: 1767–1779.PubMedCrossRefGoogle Scholar
  7. Higuchi H and Takemori S (1989) Butanedione monoxime suppresses contraction and atpase activity of rabbit skeletal-muscle. J Biochem (Tokyo) 105: 638–643.Google Scholar
  8. Hu A, Wang F and Sellers JR (2001) Mutations in human nonmuscle myosin IIA found in patients with May-Hegglin anomaly and Fechtner syndrome result in impaired enzymatic function. J Biol Chem 277: 46512–46517.CrossRefGoogle Scholar
  9. Kieke MC and Titus MA (2003) The myosin superfamily: an overview. In: Schliwa M (ed.) Molecular Motors. (pp. 3–44) Wiley-VCH, Weinheim.Google Scholar
  10. Klee CB (1977) Conformational transition accompanying the binding of Ca2+ to the protein activator of 3¢,5¢-cyclic adenosine monophosphate phosphodiesterase. Biochemistry 16: 1017–1024.PubMedCrossRefGoogle Scholar
  11. Konhilas JP and Leinwand LA (2003) Myosin myopathies. In: Schliwa M (ed.) Molecular Motors. (pp. 473–495) Wiley-VCH, Weinheim.Google Scholar
  12. Kovacs M, Wang F, Hu A, Zhang Y and Sellers JR (2003) Functional divergence of human cytoplasmic myosin II: kinetic characterization of the non-muscle IIA isoform. J Biol Chem, (in press).Google Scholar
  13. Morano I, Chai GX, Baltas LG, Lamounier-Zepter V, Lutsch G, Kott M, Haase H and Bader M (2000) Smooth-muscle contraction without smooth-muscle myosin. Nat Cell Biol 2: 371–375.PubMedCrossRefGoogle Scholar
  14. Ostap EM (2002) 2,3-Butanedione monoxime (BDM) as a myosin inhibitor. J Musc Res Cell Motility 23: 305–308.CrossRefGoogle Scholar
  15. Pato MD, Sellers JR, Preston YA, Harvey EV and Adelstein RS (1996) Baculovirus expression of chicken nonmuscle heavy meromyosin II-B – Characterization of alternatively spliced isoforms. J Biol Chem 271: 2689–2695.PubMedCrossRefGoogle Scholar
  16. Sakamoto T, Wang F, Schmitz S, Xu YH, Xu Q, Molloy JE, Veigel C and Sellers JR (2003) Neck length and processivity of myosin V. J Biol Chem 278: 29201–29207.PubMedCrossRefGoogle Scholar
  17. Sellers JR (1985) Mechanism of the phosphorylation-dependent regulation of smooth muscle heavy meromyosin. J Biol Chem 260: 15815–15819.PubMedGoogle Scholar
  18. Sellers JR (1999) Myosins. Oxford University Press, Oxford.Google Scholar
  19. Sellers JR, Cuda G, Wang F and Homsher E (1993) Myosin-specific adaptations of motility assays. In: Scholey JM (ed.) Motility Assays for Motor Proteins (pp. 23–49), Academic Press, Inc., San Diego, CA, USA.Google Scholar
  20. Sellin LC and Mcardle JJ (1994) Multiple effects of 2,3-butanedione monoxime. Pharmacology & Toxicology 74: 305–313.CrossRefGoogle Scholar
  21. Spudich JA and Watt S (1971) The regulation of rabbit skeletal muscle contraction. J Biol Chem 246: 4866–4871.PubMedGoogle Scholar
  22. Straight AF, Cheung A, Limouze J, Chen I, Westwood NJ, Sellers JR and Mitchison TJ (2003) Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor. Science 299: 1743–1747.PubMedCrossRefGoogle Scholar
  23. Wang F, Chen L, Arcucci O, Harvey EV, Bowers B, Xu Y, Hammer JA III and Sellers JR (2000) Effect of ADP and ionic strength on the kinetic and motile properties of recombinant mouse myosin V. J Biol Chem 275: 4329–4335.PubMedCrossRefGoogle Scholar
  24. Wang F, Kovacs M, Hu AH, Limouze J, Harvey EV and Sellers JR (2003) Kinetic mechanism of non-muscle myosin IIB – Functional adaptations for tension generation and maintenance. J Biol Chem 278: 27439-27448PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • John Limouze
    • 1
  • Aaron F. Straight
    • 2
  • Timothy Mitchison
    • 2
  • James R. Sellers
    • 1
  1. 1.Laboratory of Molecular Cardiology, National Heart, Lung and Blood InstituteNational Institutes of HealthMD
  2. 2.Institute for Chemistry and Cell Biology, Harvard Medical SchoolMA

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