Back to Search
Start Over
High-Affinity Functional Fluorescent Ligands for Human β-Adrenoceptors
- Source :
- Scientific Reports, Scientific Reports, Vol 7, Iss 1, Pp 1-15 (2017)
- Publication Year :
- 2017
- Publisher :
- Springer Science and Business Media LLC, 2017.
-
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.
- Subjects :
- 0301 basic medicine
Agonist
Intravital Microscopy
medicine.drug_class
Confocal
lcsh:Medicine
Biology
Ligands
Article
03 medical and health sciences
Cell Line, Tumor
Fluorescence Resonance Energy Transfer
medicine
Humans
lcsh:Science
Receptor
Fluorescent Dyes
G protein-coupled receptor
Microscopy, Confocal
Multidisciplinary
Ligand binding assay
lcsh:R
STED microscopy
Adrenergic beta-Agonists
Fluorescence
Molecular biology
Molecular Imaging
HEK293 Cells
030104 developmental biology
Förster resonance energy transfer
Molecular Probes
Biophysics
lcsh:Q
Receptors, Adrenergic, beta-2
Receptors, Adrenergic, beta-1
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 7
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....15db9a814db1cc86e9a9245f1a54453c