Humpolickova Jana, Mejdrová Ivana, Matousova Marika, Nencka Radim, Boura Evzen
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i , Flemingovo nám. 2, 166 10 Prague 6, Czech Republic.
Department of Chemistry of Natural Compounds, Institute of Chemical Technology Prague , Technicka 5, Prague 166 28, Czech Republic.
J Med Chem. 2017 Jan 12;60(1):119-127. doi: 10.1021/acs.jmedchem.6b01466. Epub 2016 Dec 22.
The lipid kinase phosphatidylinositol 4-kinase IIIβ (PI4KB) is an essential host factor for many positive-sense single-stranded RNA (+RNA) viruses including human pathogens hepatitis C virus (HCV), Severe acute respiratory syndrome (SARS), coxsackie viruses, and rhinoviruses. Inhibitors of PI4KB are considered to be potential broad-spectrum virostatics, and it is therefore critical to develop a biochemical understanding of the kinase. Here, we present highly potent and selective fluorescent inhibitors that we show to be useful chemical biology tools especially in determination of dissociation constants. Moreover, we show that the coumarin-labeled inhibitor can be used to image PI4KB in cells using fluorescence-lifetime imaging microscopy (FLIM) microscopy.
脂质激酶磷脂酰肌醇4激酶IIIβ（PI4KB）是许多正链单链RNA（+RNA）病毒的重要宿主因子，这些病毒包括人类病原体丙型肝炎病毒（HCV）、严重急性呼吸综合征（SARS）、柯萨奇病毒和鼻病毒。PI4KB抑制剂被认为是潜在的广谱病毒抑制剂，因此深入了解该激酶的生化特性至关重要。在此，我们展示了高效且选择性的荧光抑制剂，这些抑制剂是有用的化学生物学工具，尤其可用于测定解离常数。此外，我们表明香豆素标记的抑制剂可用于通过荧光寿命成像显微镜（FLIM）在细胞中对PI4KB进行成像。
