title: Fluorescent Inhibitors as Tools To Characterize Enzymes: Case Study of the Lipid Kinase Phosphatidylinositol 4-Kinase IIIβ (PI4KB).
abstract: 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.
authors.affiliation: 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.
authors.forename: Jana
authors.initials: J
authors.lastname: Humpolickova
authors.affiliation: 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.
authors.forename: Ivana
authors.initials: I
authors.lastname: Mejdrová
authors.affiliation: 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.
authors.forename: Marika
authors.initials: M
authors.lastname: Matousova
authors.affiliation: 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.
authors.forename: Radim
authors.identifier: 0000-0001-6167-0380
authors.initials: R
authors.lastname: Nencka
authors.affiliation: 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.
authors.forename: Evzen
authors.identifier: 0000-0002-9652-4065
authors.initials: E
authors.lastname: Boura
mesh_terms: D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D062105:Molecular Docking Simulation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D017853:Phosphotransferases (Alcohol Group Acceptor); C581087:PI4KB protein, human