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Sci Rep. 2016 May 31;6:26141. doi: 10.1038/srep26141.

A highly soluble, non-phototoxic, non-fluorescent blebbistatin derivative.

Author information

1
Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
2
Printnet Ltd., H-2212 Veresegyház, Hungary.
3
Department of Genetics, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
4
Brain Plasticity Unit, ESPCI-Paris-Tech, CNRS UMR8249, Paris, France.
5
MTA-ELTE Molecular Biophysics Research Group, Department of Biochemistry, Eötvös Loránd University, H-1117 Budapest, Hungary.
6
Optopharma Ltd., H-1015 Budapest, Hungary.

Abstract

Blebbistatin is a commonly used molecular tool for the specific inhibition of various myosin II isoforms both in vitro and in vivo. Despite its popularity, the use of blebbistatin is hindered by its poor water-solubility (below 10 micromolar in aqueous buffer) and blue-light sensitivity, resulting in the photoconversion of the molecule, causing severe cellular phototoxicity in addition to its cytotoxicity. Furthermore, blebbistatin forms insoluble aggregates in water-based media above 10 micromolar with extremely high fluorescence and also high adherence to different types of surfaces, which biases its experimental usage. Here, we report a highly soluble (440 micromolar in aqueous buffer), non-fluorescent and photostable C15 amino-substituted derivative of blebbistatin, called para-aminoblebbistatin. Importantly, it is neither photo- nor cytotoxic, as demonstrated on HeLa cells and zebrafish embryos. Additionally, para-aminoblebbistatin bears similar myosin II inhibitory properties to blebbistatin or para-nitroblebbistatin (not to be confused with the C7 substituted nitroblebbistatin), tested on rabbit skeletal muscle myosin S1 and on M2 and HeLa cells. Due to its drastically improved solubility and photochemical feature, as well as lack of photo- or cytotoxicity, para-aminoblebbistatin may become a feasible replacement for blebbistatin, especially at applications when high concentrations of the inhibitor or blue light irradiation is required.

PMID:
27241904
PMCID:
PMC4886532
DOI:
10.1038/srep26141
[Indexed for MEDLINE]
Free PMC Article

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