Conserved residues in the extracellular loop 2 regulate Stachel-mediated activation of ADGRG2
MetadataShow full item record
AbstractCleavage 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.
extracellular loop 2 (ECL2)
Identifier to cite or link to this itemhttp://hdl.handle.net/10713/16160
- Optimization of a peptide ligand for the adhesion GPCR ADGRG2 provides a potent tool to explore receptor biology.
- Authors: Sun Y, Zhang D, Ma ML, Lin H, Song Y, Wang J, Ma C, Yu K, An W, Guo S, He D, Yang Z, Xiao P, Hou G, Yu X, Sun JP
- Issue date: 2021 Jan-Jun
- Activation of Adhesion G Protein-coupled Receptors: AGONIST SPECIFICITY OF STACHEL SEQUENCE-DERIVED PEPTIDES.
- Authors: Demberg LM, Winkler J, Wilde C, Simon KU, Schön J, Rothemund S, Schöneberg T, Prömel S, Liebscher I
- Issue date: 2017 Mar 17
- Tethered peptide activation mechanism of the adhesion GPCRs ADGRG2 and ADGRG4.
- Authors: Xiao P, Guo S, Wen X, He QT, Lin H, Huang SM, Gou L, Zhang C, Yang Z, Zhong YN, Yang CC, Li Y, Gong Z, Tao XN, Yang ZS, Lu Y, Li SL, He JY, Wang C, Zhang L, Kong L, Sun JP, Yu X
- Issue date: 2022 Apr
- 7TM Domain Structure of Adhesion GPCRs.
- Authors: de Graaf C, Nijmeijer S, Wolf S, Ernst OP
- Issue date: 2016
- <i>Stachel</i>-independent modulation of GPR56/ADGRG1 signaling by synthetic ligands directed to its extracellular region.
- Authors: Salzman GS, Zhang S, Gupta A, Koide A, Koide S, Araç D
- Issue date: 2017 Sep 19