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Quantitative analysis of fluorescent ligand binding to dopamine D 3 receptors using live‐cell microscopy
Author(s) -
Allikalt Anni,
Laasfeld Tõnis,
Ilisson Mihkel,
Kopanchuk Sergei,
Rinken Ago
Publication year - 2021
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.15519
Subject(s) - ligand (biochemistry) , receptor , ligand binding assay , dopamine receptor , biophysics , fluorescence microscope , g protein coupled receptor , fluorophore , fluorescence , biology , chemistry , microbiology and biotechnology , biochemistry , physics , quantum mechanics
We implemented an assay combining fluorescence and bright‐field microscopy to measure ligand binding to dopamine D 3 receptors in live mammalian cells. For fluorescence intensity quantification from microscopy images, we developed a machine learning‐based user‐friendly software membrane tools . For the experiments, a novel fluorescent ligand NAPS‐Cy3B was synthesized. The subnanomolar affinity of NAPS‐Cy3B makes it a suitable ligand for the characterization of D 3 receptors and its ligands in live HEK293 cells.