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Specificity of blebbistatin, an inhibitor of myosin II

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Abstract

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.

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REFERENCES

  • Adelstein RS and Klee CB (1981) Purification and characterization of smooth muscle myosin light chain kinase. J Biol Chem 256: 7501–7509.

    PubMed  CAS  Google Scholar 

  • Avraham KB (2003) Molecular motors in sensory defects. In: Schliwa M. (ed.) Molecular Motors. (pp. 511–537) Wiley-VCH, Wenheim.

    Google Scholar 

  • Berg JS, Powell BC and Cheney RE (2001) A millennial myosin census. Mol Biol Cell 12: 780–794.

    PubMed  CAS  Google Scholar 

  • Brzeska H and Korn ED (1996) Regulation of class I and class II myosins by heavy chain phosphorylation. J Biol Chem 271: 16983–16986.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Higuchi H and Takemori S (1989) Butanedione monoxime suppresses contraction and atpase activity of rabbit skeletal-muscle. J Biochem (Tokyo) 105: 638–643.

    CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Konhilas JP and Leinwand LA (2003) Myosin myopathies. In: Schliwa M (ed.) Molecular Motors. (pp. 473–495) Wiley-VCH, Weinheim.

    Google Scholar 

  • 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).

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Ostap EM (2002) 2,3-Butanedione monoxime (BDM) as a myosin inhibitor. J Musc Res Cell Motility 23: 305–308.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Sellers JR (1985) Mechanism of the phosphorylation-dependent regulation of smooth muscle heavy meromyosin. J Biol Chem 260: 15815–15819.

    PubMed  CAS  Google Scholar 

  • Sellers JR (1999) Myosins. Oxford University Press, Oxford.

    Google Scholar 

  • 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 

  • Sellin LC and Mcardle JJ (1994) Multiple effects of 2,3-butanedione monoxime. Pharmacology & Toxicology 74: 305–313.

    Article  CAS  Google Scholar 

  • Spudich JA and Watt S (1971) The regulation of rabbit skeletal muscle contraction. J Biol Chem 246: 4866–4871.

    PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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: 27439-27448

    Article  PubMed  CAS  Google Scholar 

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Limouze, J., Straight, A.F., Mitchison, T. et al. Specificity of blebbistatin, an inhibitor of myosin II. J Muscle Res Cell Motil 25, 337–341 (2004). https://doi.org/10.1007/s10974-004-6060-7

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  • DOI: https://doi.org/10.1007/s10974-004-6060-7

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