Endothelin GPCR Assays

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Background of Endothelin Receptors

Endothelin is a natural ligand of two G protein-coupled receptors, endothelin_A and endothelin_B receptors (ET_A and ET_B). Endothelin is an endogenous vasoconstricting peptide, which plays a role in cell proliferation, vascular tone, vascular homeostasis, and neuronal function. The overexpression of endothelin leads to hypertension, heart disease, and other related diseases.

Fig.1. Crystal structure of endothelin ETB receptor bound to bosentan. (Davenport, 2019)

Distribution and Function of Endothelin Receptors

ET receptors are broadly located in all tissues, particularly in blood vessels, such as vascular smooth muscle cells and endothelial cells on the vessel wall, and non-vascular tissues, including epithelial cells, astrocytes, and neurons. ET receptor antagonists have been already demonstrated that can be used for the treatment of pulmonary arterial hypertension, while the ET_B receptor agonists are effective for chemotherapy, neuroprotection, renal failure, and reducing proteinuria in diabetic nephropathy. However, ET_A receptor agonists have not been explored to date. ET_B monoclonal antibodies and the ET_B receptor agonists and antagonists are developing with advanced technologies as new pharmacological options for the future.

Subtypes and Mechanism of Endothelin Receptors

Endothelin receptors are a family of G protein-coupled receptors, including ET_A and ET_B. ET_A receptors couple G_q/G₁₁ and ET_B receptors facilitate signal transduction through G_s, G_i/G_o, and G_q/G₁₁ proteins.

Receptor	Gene	Mechanism	agonists	Antagonists
ET_A receptor	EDNRA	ET_A receptor binds to the ligand and G_q/G₁₁ protein to stimulate signal transduction ET_A receptor activates phospholipase C, phospholipase A₂, and phospholipase D ET_A receptor regulates calcium signaling, which responds to ET-1, inducing vasoconstriction via the generation of IP3 and DAG and facilitates cell proliferation	ET-1 ET-2 sarafotoxin S6b	atrasentan
ET_B receptor	EDNRB	ET_B receptor binds to theligands, G_s protein, G_i/G_o protein and G_q/G₁₁ protein to promote signal transduction ET_B receptor activates phospholipase C, phospholipase A₂, and phospholipase D ET_B receptor increases tyrosine phosphorylation of cellular proteins, stimulating MAPK, and promotes DNA synthesis in certain cells like astrocytes	ET-3 sarafotoxin S6b IRL1620 ET-1 BQ 3020	A192621 BQ788 IRL 2500 bosentan K-8794 RES7011

Assay List of Endothelin Receptors

Creative Biolabs can provide a range of assays of endothelin receptors. You can choose the assay in the list or contact us for more information:

EDNRA EDNRB

Assay No.	Assay Name	Host Cell	Assay Type
Calcium Flux Assay
S01YF-0722-KX105	Magic™ Human EDNRA In Vitro Calcium Assay, HEK293	HEK293	Calcium Flux Assay
IP1 Assay
S01YF-0722-KX20	Magic™ Human EDNRA In Vitro Agonist & Antagonist IP1 Assay, CHO	CHO	IP1 Assay

Assay No.	Assay Name	Host Cell	Assay Type	Datasheet
Calcium Flux Assay
S01YF-0722-KX106	Magic™ Human EDNRB In Vitro Calcium Assay, HEK293	HEK293	Calcium Flux Assay

Published Data

Paper Title	G protein- and β-arrestin signaling profiles of endothelin derivatives at the type A endothelin receptor
Journal	Kidney360
Published	2021
Abstract	The type A endothelin receptor (ET_AR) is a GPCR with a peptide agonist that couples primarily through G_q and G_i and promotes vasoconstriction. Antagonists of the ET_AR have shown promising results in vasoconstriction action. However, side effects limit widespread use. Endothelin-1 (ET-1) is a potent vasoconstrictor in the cardiovascular system via mediating the ET_AR. The study aimed to investigate if there was any significant ligand bias between endothelin derivatives, and whether the derivatives have any physiologic effects on the cardiovascular system. G protein signaling assay and β-arrestin 2 recruitment assay at the ET_AR were applied for the study. In addition, the effects of ET-1 on the vasopressor response in wild-type and β-arrestin 1 and 2 KO mice were tested. The physiologic characteristics of ET_AR-biased agonists may have the potential to be tool chemicals or novel therapies.
Result	The study has demonstrated that the endothelins activated a wide range of G proteins at the ET_AR. However, the results have shown no significant agonistic effect of the endothelin derivatives on different G proteins. Regarding their levels of agonism, endothelin derivatives did show relationships between structure and activity. The chronic responses to ET-1 did not differ between β-arrestin 2 knockout mice and wild-type mice in terms of blood pressure, while the acute pressor reaction to ET-1 did not differ between β-arrestin 1 and 2 knockout mice. G proteins were responsible for the vasoconstriction, and the ET_AR did not have considerable desensitization from β-arrestins. These results implied that G protein- and β-arrestin-biased ET_AR agonists would differ physiologically from balanced agonists, even if the endothelin peptide scaffold did not seem appropriate for creating such ligands. Agonists and antagonists targeting ET_AR should use nonpeptide scaffolds to lead to biased signaling, which may represent a novel way to block many of the deleterious actions of endothelins, while perhaps minimizing significant side effects. Fig.2. Concentration response of endothelin-1 (ET-1), ET-2, and ET-3 in assays of G protein signaling and β-arrestin recruitment. (Xiong, 2021)

References

Davenport A.P.; et al. Endothelin receptors (version 2019.4) in the IUPHAR/BPS Guide to Pharmacology Database. IUPHAR/BPS Guide to Pharmacology CITE. 2019, 2019(4).
Xiong, X.; et al. G protein-and β-arrestin signaling profiles of endothelin derivatives at the type a endothelin receptor. Kidney360. 2021, 2(7): 1124.

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