title: HTS-compatible FRET-based conformational sensors clarify membrane receptor activation.
abstract: Cell surface receptors represent a vast majority of drug targets. Efforts have been conducted to develop biosensors reporting their conformational changes in live cells for pharmacological and functional studies. Although Förster resonance energy transfer (FRET) appears to be an ideal approach, its use is limited by the low signal-to-noise ratio. Here we report a toolbox composed of a combination of labeling technologies, specific fluorophores compatible with time-resolved FRET and a novel method to quantify signals. This approach enables the development of receptor biosensors with a large signal-to-noise ratio. We illustrate the usefulness of this toolbox through the development of biosensors for various G-protein-coupled receptors and receptor tyrosine kinases. These receptors include mGlu, GABAB, LH, PTH, EGF and insulin receptors among others. These biosensors can be used for high-throughput studies and also revealed new information on the activation process of these receptors in their cellular environment.
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.|Cisbio Bioassays, Codolet, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Scholler
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: David
authors.initials: D
authors.lastname: Moreno-Delgado
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Lecat-Guillet
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Doumazane
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: Carine
authors.initials: C
authors.lastname: Monnier
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Charrier-Savournin
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.|Cisbio Bioassays, Codolet, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Fabre
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Chouvet
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Soldevila
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Lamarque
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Donsimoni
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Roux
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Jurriaan M
authors.identifier: 0000-0001-9017-5856
authors.initials: JM
authors.lastname: Zwier
authors.affiliation: Cisbio Bioassays, Codolet, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Trinquet
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Rondard
authors.affiliation: Institut de Génomique Fonctionnelle, (IGF), CNRS, INSERM, Université de Montpellier, Montpellier, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Pin
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*
chemical_list: D043562:Receptors, G-Protein-Coupled; D020794:Receptor Protein-Tyrosine Kinases