High-Affinity Functional Fluorescent Ligands for Human β-Adrenoceptors

Scientific Reports
Gyuzel Yu MitronovaStefan W Hell

Abstract

Visualization of the G-protein coupled receptor (GPCR) is of great importance for studying its function in a native cell. We have synthesized a series of red-emitting fluorescent probes targeting β-adrenergic receptor (βAR) that are compatible with confocal and Stimulated Emission Depletion (STED) microscopy as well as with Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) binding assay in living cells. The probe based on the agonist BI-167107 and fluorescent dye KK114 demonstrates nanomolar binding affinity and up to nine-fold β2AR selectivity over β1AR. Carazolol-derived probes are fluorogenic and allow no-wash imaging experiments. STED microscopy of β2ARs stained at the native expression level on pancreatic CAPAN cells provides two-fold improvement in lateral optical resolution over confocal mode and reveals the formation of receptor microdomains. These probes retain their functional (agonist or antagonist) properties, allowing simultaneous modulation of cyclic adenosine monophosphate (cAMP) levels and receptor internalization as well as imaging receptor localization.

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Citations

Sep 26, 2018·Expert Opinion on Drug Discovery·Christos Iliopoulos-TsoutsouvasSpyros P Nikas

Methods Mentioned

BETA
superresolution microscopy
FRET
biosensor
saturation
Fluorescence
transfection
saturation binding assay

Related Concepts

Cyclic AMP
Beta-2 Adrenergic Receptors
Diagnostic Imaging
Fluorescent Dyes
Fluorescent Probes
Fluorogenic Substrate
Structure of Alpha Cell of Islet
Positron-Emission Tomography
Beta-adrenergic receptor
Penicillin-Binding Proteins

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