title: A highly soluble, non-phototoxic, non-fluorescent blebbistatin derivative.
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.
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.|Printnet Ltd., H-2212 Veresegyház, Hungary.
authors.forename: Boglárka H
authors.initials: BH
authors.lastname: Várkuti
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.|Printnet Ltd., H-2212 Veresegyház, Hungary.
authors.forename: Miklós
authors.initials: M
authors.lastname: Képiró
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: István Ádám
authors.initials: IÁ
authors.lastname: Horváth
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: László
authors.initials: L
authors.lastname: Végner
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: Szilvia
authors.initials: S
authors.lastname: Ráti
authors.affiliation: Department of Genetics, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: Áron
authors.initials: Á
authors.lastname: Zsigmond
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: György
authors.initials: G
authors.lastname: Hegyi
authors.affiliation: Brain Plasticity Unit, ESPCI-Paris-Tech, CNRS UMR8249, Paris, France.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Lenkei
authors.affiliation: Department of Genetics, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.
authors.forename: Máté
authors.initials: M
authors.lastname: Varga
authors.affiliation: Department of Biochemistry, Institute of Biology, Eötvös Loránd University, H-1117 Budapest, Hungary.|MTA-ELTE Molecular Biophysics Research Group, Department of Biochemistry, Eötvös Loránd University, H-1117 Budapest, Hungary.|Optopharma Ltd., H-1015 Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Málnási-Csizmadia
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D011232:Chemical Precipitation; D017484:Dermatitis, Phototoxic; D004355:Drug Stability; D005453:Fluorescence; D006367:HeLa Cells; D006576:Heterocyclic Compounds, 4 or More Rings / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology / Q000633:toxicity*; D006801:Humans; D024462:Myosin Type II / Q000037:antagonists & inhibitors; D011817:Rabbits; D012995:Solubility; D016019:Survival Analysis; D015027:Zebrafish
chemical_list: D006576:Heterocyclic Compounds, 4 or More Rings; C472645:blebbistatin; D024462:Myosin Type II