β-arrestin2 recruitment by β-adrenergic receptor agonists and antagonists.
- Author:
Yi-Ran WANG
1
;
De-Qin CHENG
2
;
Lan MA
1
;
Xing LIU
3
Author Information
1. Pharmacology Research Center, Shanghai Medical College, Fudan University, Shanghai 200032, China.
2. Institutes of Brain Science, Fudan University, Shanghai 200032, China.
3. Pharmacology Research Center, Shanghai Medical College, Fudan University, Shanghai 200032, China. xingliu@fudan.edu.cn.
- Publication Type:Journal Article
- MeSH:
Humans;
beta-Arrestin 2/metabolism*;
HEK293 Cells;
Adrenergic beta-Agonists/pharmacology*;
Isoproterenol/pharmacology*;
Receptors, Adrenergic, beta-2/metabolism*;
Norepinephrine/pharmacology*
- From:
Acta Physiologica Sinica
2022;74(6):993-1004
- CountryChina
- Language:Chinese
-
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.
