(-)-Blebbistatin
(-)-Blebbistatin (CAS 856925-71-8) is a cell-permeable small molecule inhibitor targeting non-muscle myosin II (NM II), an actin-dependent motor protein involved in cell adhesion, migration, and differentiation. It selectively binds the myosin-ADP-phosphate complex, slowing phosphate release and thereby suppressing Mg-ATPase and contractile activities mediated by actomyosin interactions. It exhibits inhibitory potency in various model systems (IC50: 0.5–5.0 μM), with minimal effects on myosin isoforms I, V, X, and reduced activity toward smooth muscle myosin II (IC50 ~80 μM). Its reversible modulation of NM II renders it useful in research on cytoskeletal dynamics, cell mechanics, and related pathophysiological conditions.
References:
[1]. Straight AF, Cheung A, Limouze J, et al. Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor. Science, 2003, 299:1743–1747.
[2]. Kovacs M, Toth J, Hetenyi C, Malnasi-Csizmadia A, Sellers JR. Mechanism of blebbistatin inhibition of myosin II. J Biol Chem, 2004, 279:35557–35563.
[3]. Limouze J, Straight AF, Mitchison T, Sellers JR. Specificity of blebbistatin, an inhibitor of myosin II. J Muscle Res Cell Motil, 2004, 25:337–341.
[4]. Miguel Vicente-Manzanares, Xuefei Ma, Robert S. Adelstein,Alan Rick Horwitz. Non-muscle myosin II takes centre stage in cell adhesion and migration.Nat Rev Mol Cell Biol, 2009 Nov, 10(11): 778–790.
[5]. Burt RA, Joseph JE, Milliken S, Collinge JE, Kile BT. Description of a novel mutation leading to MYH9-related disease. Thrombosis Research, 2008, 122(6): 861-863.
[6]. Butcher DT, Alliston T, Weaver VM. A tense situation: forcing tumour progression. Nature Reviews Cancer, 2009 Feb, 9(2):108-122.
[7]. Chen Y, Boukour S, Milloud R, Favier R, Saposnik B, Schlegel N, et al. The abnormal proplatelet formation in MYH9-related macrothrombocytopenia results from an increased actomyosin contractility and is rescued by myosin IIA inhibition. Journal of thrombosis and haemostasis : JTH , 2013, 11:2163-2175.
[8]. Fedorov VV, Lozinsky IT, Sosunov EA, Anyukhovsky EP, Rosen MR, Balke CW, et al. Application of blebbistatin as an excitation-contraction uncoupler for electrophysiologic study of rat and rabbit hearts. Heart rhythm: the official journal of the Heart Rhythm Society, 2007, 4:619-626.
[9]. Zhang X-h, Aydin M, Kuppam D, Melman A, DiSanto ME. In Vitro and In Vivo Relaxation of Corpus Cavernosum Smooth Muscle by the Selective Myosin II Inhibitor, Blebbistatin. The Journal of Sexual Medicine, 2009, 6:2661-2671.
[10]. Zhang M, Rao PV. Blebbistatin, a novel inhibitor of myosin II ATPase activity, increases aqueous humor outflow facility in perfused enucleated porcine eyes. Investigative ophthalmology & visual science, 2005, 46:4130-4138.
[11]. Lucas-Lopez C, Allingham JS, Lebl T, Lawson CP, Brenk R, Sellers JR, et al. The small molecule tool (S)-(-)-blebbistatin: novel insights of relevance to myosin inhibitor design. Organic & biomolecular chemistry, 2008, 6:2076-2084.
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| Physical Appearance | A solid |
| Storage | Store at -20°C |
| M.Wt | 292.33 |
| Cas No. | 856925-71-8 |
| Formula | C18H16N2O2 |
| Solubility | insoluble in EtOH; insoluble in H2O; ≥14.62 mg/mL in DMSO |
| Chemical Name | (S)-3a-hydroxy-6-methyl-1-phenyl-3,3a-dihydro-1H-pyrrolo[2,3-b]quinolin-4(2H)-one |
| SDF | Download SDF |
| Canonical SMILES | O[C@@]12CCN(C3=CC=CC=C3)C1=NC4=CC=C(C)C=C4C2=O |
| Shipping Condition | Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice. |
| General tips | We do not recommend long-term storage for the solution, please use it up soon. |
| Kinase experiment [1]: | |
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Actin-activated MgATPase assay |
Actin-activated MgATPase activity is measured using an NADH-coupled assay in a Beckman DU 640 spectrophotometer. Blebbistatin [a racemic mixture of the (+) and (-) enantiomers] is added from stocks dissolved in DMSO and the DMSO concentration is maintained at a constant concentration of 5% in all samples. |
| Cell experiment [2, 3]: | |
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Cell lines |
Bovine corneal endothelial cells (BCECs) |
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Preparation method |
The solubility of this compound in DMSO is > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
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Reacting condition |
10 μM for 5 min; or 5 μM for 20 minutes |
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Applications |
Blebbistatin effectively inhibited actin-myosin interaction in mouse cardiac muscle through a mechanism independent on Ca2+ influx and activation systems. In addition, blebbistatin prevented thrombin-mediated inhibition of intercellular calcium wave propagation in bovine corneal endothelial cells. |
| Animal experiment [4]: | |
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Animal models |
Zebrafish embryos model |
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Dosage form |
5-25 μM |
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Applications |
Blebbistatin treatment induced zebrafish embryos cardia bifida in a dose-dependent manner. |
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Other notes |
Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal. |
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References: 1. Limouze, J., Straight, A. F., Mitchison, T. and Sellers, J. R. (2004) Specificity of blebbistatin, an inhibitor of myosin II. J Muscle Res Cell Motil. 25, 337-341 2. Ponsaerts, R., D'Hondt, C., Bultynck, G., Srinivas, S. P., Vereecke, J. and Himpens, B. (2008) The myosin II ATPase inhibitor blebbistatin prevents thrombin-induced inhibition of intercellular calcium wave propagation in corneal endothelial cells. Invest Ophthalmol Vis Sci. 49, 4816-4827 3. Dou, Y., Arlock, P. and Arner, A. (2007) Blebbistatin specifically inhibits actin-myosin interaction in mouse cardiac muscle. Am J Physiol Cell Physiol. 293, C1148-1153 4. Wang, X., Chong, M., Wang, H., Zhang, J., Xu, H. and Liu, D. (2015) Block the function of nonmuscle myosin II by blebbistatin induces zebrafish embryo cardia bifida. In Vitro Cell Dev Biol Anim. 51, 211-217 |
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Quality Control & MSDS
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