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β-arrestin2 recruitment by β-adrenergic receptor agonists and antagonists 

WANG Yi-Ran¹, CHENG De-Qin², MA Lan¹, LIU Xing^1,*

¹ Pharmacology Research Center, Shanghai Medical College, Fudan University, Shanghai 200032, China；²Institutes of Brain Science, Fudan University, Shanghai 200032, China

Abstract

A large number of β-adrenergic receptor (β-AR) agonists and antagonists are widely used in the treatment of cardiovascular diseases and other diseases. Nonetheless, it remains unclear whether these commonly used β-AR drugs can activate downstream β- arrestin-biased signaling pathways. The objective of this study was to investigate β-arrestin2 recruitment effects of β-AR agonists and antagonists that were commonly used in clinical practice. We used TANGO (transcriptional activation following arrestin translocation) assay to detect the β-arrestin2 recruitment by β-AR ligands in HEK293 cell line (HTLA cells) stably transfected with tetracycline transactivator protein (tTA) dependent luciferase reporter and β-arrestin2-TEV fusion gene. Upon activation of β-AR by a β-AR ligand, β-arrestin2 was recruited to the C terminus of the receptor, followed by cleavage of the G protein-coupled receptors (GPCRs) fusion protein at the TEV protease-cleavage site. The cleavage resulted in the release of tTA, which, after being transported to the nucleus, activated transcription of the luciferase reporter gene. The results showed that β-AR non-selective agonists epinephrine, noradrenaline and isoprenaline all promoted β-arrestin2 recruitment at β1-AR and β2-AR. β1-AR selective agonists dobutamine and denopamine both promoted β-arrestin2 recruitment at β1-AR. β2-AR selective agonists procaterol and salbutamol promoted β-arrestin2 recruitment at β2-AR. β-AR non-selective antagonists alprenolol and pindolol promoted β-arrestin2 recruitment at β1-AR. β1-AR selective antagonists celiprolol and bevantolol showed β-arrestin2 recruitment at β1-AR. β2-AR selective antagonists butoxamine showed β-arrestin2 recruitment at β1-AR. These results provide some clues for the potential action of β-AR drugs, and lay a foundation for the screening of β-arrestin-biased β-AR ligands.

Key words: β-adrenergic receptors; β-arrestin2; G protein-coupled receptors; luciferase reporter gene

Received: 　　Accepted:

Corresponding author: 刘星　　E-mail: xingliu@fudan.edu.cn

Citing This Article：

WANG Yi-Ran, CHENG De-Qin, MA Lan, LIU Xing. β-arrestin2 recruitment by β-adrenergic receptor agonists and antagonists . Acta Physiol Sin 2022; 74 (6): 993-1004 (in Chinese with English abstract).