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(-)-Blebbistatin

现货
Catalog No.
B1387
非肌肉肌球蛋白II型ATP酶抑制剂
组合的产品项目
规格价格库存 数量
10mM (in 1mL DMSO)
¥ 650.00
现货
10mg
¥ 1,190.00
现货
25mg
¥ 2,590.00
现货

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Background

(-)-Blebbistatin is a cell-permeable non-muscle myosin II ATPases inhibitor with an IC50 range of 2 μM [1,2].

Non-muscle myosin II (NM II), an actin-binding protein, plays a central role in regulation of cell migration, adhesion and differentiation [4]. Recent insight into the importance of NM II in these processes has been highlighted by genetic deletion and mutation methods that discovered NM II mutations affect the function of a wide range of proteins and cause monogenic diseases [5.6].

(-)-Blebbistatin is a small molecule inhibitor and preferentially binds to the myosin-ADP-Pi complex to slow down phosphate release [2]. The inhibitor completely eliminate contraction of activity of actin-activated Mg-ATPase and motility of myosins II for several species in vitro (IC50 = 0.5-5.0 μM) [8,9], but it has poor effects on smooth muscle myosin II (IC50 =80 μM) and myosins I,V, and X [3]. Furthermore, blebbistatin can potently inhibit mammalian arterial smooth muscle (IC50=5 μM) [9]. The property that blebbistatin blocks myosin II in an actin-detached state and prevents rigid actomyosin cross-linking is a great advantage in vivo applications [2,11].

In a constant-pressure grant perfusion model system, the CB and TM cells were treated with blebbistatin (10-200 M) and cell morphology was changed, and actin stress fiber content decreased. The blebbistatin effect was completely reversible by washout within 24 hours [10]. Blebbistatin inhibited single cellular contraction without altering the morphologies of intracellular calcium transients (IC50 = 0.43 μM). Exposure to UV light at wavelengths below 488 nm can also cause blebbistatin rapidly suppressed. [8].

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.

文献引用

1.Wang Y, Li Y, et al. "The cerebral cavernous malformation disease causing gene KRIT1 participates in intestinal epithelial barrier maintenance and regulation." FASEB J. 2018 Sep 25:fj201800343R. PMID:30252535
2.Petrova IO, Tyganov SA, et al. "Early Deсline in Rat Soleus Passive Tension with Hindlimb Unloading: Inactivation of Cross-bridges or Activation of Calpains?" Dokl Biochem Biophys. 2018 Jul;481(1):205-207. PMID:30168060
3.Chen Y, Yu Y, et al. "Bradykinin promotes migration and invasion of hepatocellular carcinoma cells through TRPM7 and MMP2." Exp Cell Res. 2016 Sep 29. PMID:27693494

Chemical Properties

Physical AppearanceA solid
StorageStore at -20°C
M.Wt292.33
Cas No.856925-71-8
FormulaC18H16N2O2
Solubility≥14.6 mg/mL in DMSO, <1.46 mg/mL in EtOH, <1.55 mg/mL in H2O
Chemical Name(S)-3a-hydroxy-6-methyl-1-phenyl-3,3a-dihydro-1H-pyrrolo[2,3-b]quinolin-4(2H)-one
SDFDownload SDF
Canonical SMILESO[C@@]12CCN(C3=CC=CC=C3)C1=NC4=CC=C(C)C=C4C2=O
运输条件试用装:蓝冰运输。 其他可选规格:常温运输或根据您的要求用蓝冰运输。
一般建议为了使其更好的溶解,请用37℃加热试管并在超声波水浴中震动片刻。不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。

试验操作

激酶实验 [1]:

肌动蛋白活化的MgATPase检测

利用NADH耦合方法,使用Beckman DU 640分光光度计测量肌动蛋白活化的MgATPase活性。Blebbistatin[(+)和(-)对映体的外消旋混合物]溶解在DMSO中,所有样品中DMSO浓度维持在恒定浓度5%。

细胞实验 [2, 3]:

细胞系

牛角膜内皮细胞

溶解方法

该化合物在DMSO中的溶解度> 10 mM。若配制更高浓度的溶液,一般步骤如下:请将试管置于37℃加热10分钟和/或将其置于超声波浴中震荡一段时间。原液于-20℃可放置数月。

反应时间

10 μM处理5 分钟; 或5 μM处理20分钟

应用

Blebbistatin有效抑制小鼠心肌内肌动蛋白与肌球蛋白的相互作用,其作用分子机制不是通过钙流及其活化体系。此外,Blebbistatin处理牛角膜内皮细胞能干预凝血酶抑制的细胞内钙流振荡。

动物实验 [4]:

动物模型

斑马鱼胚胎模型

剂量

5-25 μM

应用

Blebbistatin以剂量相关的方式诱导斑马鱼胚胎心脏二分叉。

注意事项

请于室内测试所有化合物的溶解度。实际溶解度和理论值可能略有不同,这是由实验系统的误差引起的,属于正常现象。

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

质量控制

化学结构

(-)-Blebbistatin

相关生物数据

(-)-Blebbistatin

相关生物数据

(-)-Blebbistatin

相关生物数据

(-)-Blebbistatin