title: Development of BODIPY FL vindoline as a novel and high-affinity pregnane X receptor fluorescent probe.
abstract: The pregnane X receptor (PXR) regulates the metabolism and excretion of xenobiotics and endobiotics by regulating the expression of drug-metabolizing enzymes and transporters. The unique structure of PXR allows it to bind many drugs and drug leads, possibly causing undesired drug-drug interactions. Therefore, it is crucial to evaluate whether chemicals or drugs bind to PXR. Fluorescence-based assays are preferred because of their sensitivity and nonradioactive nature. On the basis of our previously characterized 4 (BODIPY FL vinblastine), a high-affinity PXR probe, we developed 20 (BODIPY FL vindoline) and showed that it is a novel and potent PXR fluorescent probe with Kd of 256 nM in a time-resolved fluorescence resonance energy transfer (TR-FRET) binding assay with PXR. By using 20 (BODIPY FL vindoline) in the PXR TR-FRET assay, we obtained a more than 7-fold signal-to-background ratio and high signal stability (signal was stable for at least 120 min, and Z'-factor > 0.85 from 30 to 240 min). The assay can tolerate DMSO up to 2%. This assay has been used to evaluate a panel of PXR ligands for their PXR-binding affinities. The performance of 20 (BODIPY FL vindoline) in the PXR TR-FRET assay makes it an ideal PXR fluorescent probe, and the newly developed PXR TR-FRET assay with 20 (BODIPY FL vindoline) as a fluorescent probe is suitable for high-throughput screening to identify PXR-binding ligands.
authors.affiliation: Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital , 262 Danny Thomas Place, Mail Stop 1000, Memphis, Tennessee 38105, United States.
authors.forename: Wenwei
authors.initials: W
authors.lastname: Lin
authors.forename: Jiuyu
authors.initials: J
authors.lastname: Liu
authors.forename: Cynthia
authors.initials: C
authors.lastname: Jeffries
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
authors.forename: Richard E
authors.initials: RE
authors.lastname: Lee
authors.forename: Taosheng
authors.initials: T
authors.lastname: Chen
mesh_terms: D001896:Boron Compounds / Q000737:chemistry* / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D015195:Drug Design; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D000077297:Pregnane X Receptor; D017434:Protein Structure, Tertiary; D011987:Receptors, Steroid / Q000737:chemistry / Q000378:metabolism*; D014747:Vinblastine / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D005456:Fluorescent Dyes; D000077297:Pregnane X Receptor; D011987:Receptors, Steroid; C009667:vindoline; D014747:Vinblastine; D004121:Dimethyl Sulfoxide