Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2

Abstract Cleavage and dissociation of a large N-terminal fragment and the consequent unmasking of a short sequence (Stachel) remaining on the N-terminus have been proposed as mechanisms of activation of some members of the adhesion G protein-coupled receptor (aGPCR) family. However, the identity of...

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Autores principales:	Abanoub A. Gad, Pedram Azimzadeh, Nariman Balenga
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0b5f1f83714c4759b7b5a2d286ab6750
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spelling	oai:doaj.org-article:0b5f1f83714c4759b7b5a2d286ab67502021-12-02T15:39:59ZConserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG210.1038/s41598-021-93577-y2045-2322https://doaj.org/article/0b5f1f83714c4759b7b5a2d286ab67502021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93577-yhttps://doaj.org/toc/2045-2322Abstract Cleavage and dissociation of a large N-terminal fragment and the consequent unmasking of a short sequence (Stachel) remaining on the N-terminus have been proposed as mechanisms of activation of some members of the adhesion G protein-coupled receptor (aGPCR) family. However, the identity of residues that play a role in the activation of aGPCRs by the cognate Stachel remains largely unknown. Protein sequence alignments revealed a conserved stretch of residues in the extracellular loop 2 (ECL2) of all 33 members of the aGPCR family. ADGRG2, an orphan aGPCR, plays a major role in male fertility, Ewing sarcoma cell proliferation, and parathyroid cell function. We used ADGRG2 as a model aGPCR and generated mutants of the conserved residues in the ECL2 via site-directed mutagenesis. We show that tryptophan and isoleucine in the ECL2 are essential for receptor stability and surface expression in the HEK293 cells. By adjusting the receptor surface expression levels, we show that mutation of these residues of ECL2 ablates the Stachel-mediated activation of multiple signaling pathways of ADGRG2. This study provides a novel understanding of the role of the ECL2 in Stachel-mediated signaling and degradation of ADGRG2, which may lay the foundation for the rational design of therapeutics to target aGPCRs.Abanoub A. GadPedram AzimzadehNariman BalengaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Abanoub A. Gad Pedram Azimzadeh Nariman Balenga Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
description	Abstract Cleavage and dissociation of a large N-terminal fragment and the consequent unmasking of a short sequence (Stachel) remaining on the N-terminus have been proposed as mechanisms of activation of some members of the adhesion G protein-coupled receptor (aGPCR) family. However, the identity of residues that play a role in the activation of aGPCRs by the cognate Stachel remains largely unknown. Protein sequence alignments revealed a conserved stretch of residues in the extracellular loop 2 (ECL2) of all 33 members of the aGPCR family. ADGRG2, an orphan aGPCR, plays a major role in male fertility, Ewing sarcoma cell proliferation, and parathyroid cell function. We used ADGRG2 as a model aGPCR and generated mutants of the conserved residues in the ECL2 via site-directed mutagenesis. We show that tryptophan and isoleucine in the ECL2 are essential for receptor stability and surface expression in the HEK293 cells. By adjusting the receptor surface expression levels, we show that mutation of these residues of ECL2 ablates the Stachel-mediated activation of multiple signaling pathways of ADGRG2. This study provides a novel understanding of the role of the ECL2 in Stachel-mediated signaling and degradation of ADGRG2, which may lay the foundation for the rational design of therapeutics to target aGPCRs.
format	article
author	Abanoub A. Gad Pedram Azimzadeh Nariman Balenga
author_facet	Abanoub A. Gad Pedram Azimzadeh Nariman Balenga
author_sort	Abanoub A. Gad
title	Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
title_short	Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
title_full	Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
title_fullStr	Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
title_full_unstemmed	Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
title_sort	conserved residues in the extracellular loop 2 regulate stachel-mediated activation of adgrg2
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/0b5f1f83714c4759b7b5a2d286ab6750
work_keys_str_mv	AT abanoubagad conservedresiduesintheextracellularloop2regulatestachelmediatedactivationofadgrg2 AT pedramazimzadeh conservedresiduesintheextracellularloop2regulatestachelmediatedactivationofadgrg2 AT narimanbalenga conservedresiduesintheextracellularloop2regulatestachelmediatedactivationofadgrg2
_version_	1718385852976988160

Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2

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