A Non-imaging High Throughput Approach to Chemical Library Screening at the Unmodified Adenosine-A₃ Receptor in Living Cells.

ABSTRACT: Recent advances in fluorescent ligand technology have enabled the study of G protein-coupled receptors in their native environment without the need for genetic modification such as addition of N-terminal fluorescent or bioluminescent tags. Here, we have used a non-imaging plate reader (PHERAstar FS) to monitor the binding of fluorescent ligands to the human adenosine-A₃ receptor (A₃AR; CA200645 and AV039), stably expressed in CHO-K1 cells. To verify that this method was suitable for the study of other GPCRs, assays at the human adenosine-A₁ receptor, and β₁ and β₂ adrenoceptors (β₁AR and β₂AR; BODIPY-TMR-CGP-12177) were also carried out. Affinity values determined for the binding of the fluorescent ligands CA20064

SUBMITTER: Arruda MA 

PROVIDER: S-EPMC5733478 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature