Quantitative analysis of fluorescent ligand binding to dopamine D 3  receptors using live‐cell microscopy | Zendy

Allikalt Anni | Zendy; Laasfeld Tõnis | Zendy; Ilisson Mihkel | Zendy; Kopanchuk Sergei | Zendy; Rinken Ago | Zendy

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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 , fluorescence microscope , biophysics , 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.

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