Second extracellular loop of human glucagon-like peptide-1 receptor (GLP-1R) has a critical role in GLP-1 peptide binding and receptor activation


The glucagon-like peptide-1 receptor (GLP-1R) is a therapeutically important family B G protein-coupled receptor (GPCR) that is pleiotropically coupled to multiple signaling effectors and, with actions including regulation of insulin biosynthesis and secretion, is one of the key targets in the management of type II diabetes mellitus. However, there is limited understanding of the role of the receptor core in orthosteric ligand binding and biological activity. To assess involvement of the extracellular loop (ECL) 2 in ligand-receptor interactions and receptor activation, we performed alanine scanning mutagenesis of loop residues and assessed the impact on receptor expression and GLP-1(1-36)-NH 2 or GLP-1(7-36)-NH2 binding and activation of three physiologically relevant signaling pathways as follows: cAMP formation, intracellular Ca 2+ (Ca 2+ i) mobilization, and phosphorylation of extracellular signal-regulated kinases 1 and 2 (pERK1/2). Although antagonist peptide binding was unaltered, almost all mutations affected GLP-1 peptide agonist binding and/or coupling efficacy, indicating an important role in receptor activation. However, mutation of several residues displayed distinct pathway responses with respect to wild type receptor, including Arg-299 and Tyr-305, where mutation significantly enhanced both GLP-1(1-36)-NH2- and GLP-1(7-36)- NH 2-mediated signaling bias for pERK1/2. In addition, mutation of Cys-296, Trp-297, Asn-300, Asn-302, and Leu-307 significantly increased GLP-1(7-36)-NH 2-mediated signaling bias toward pERK1/2. Of all mutants studied, only mutation of Trp-306 to alanine abolished all biological activity. These data suggest a critical role of ECL2 of the GLP-1R in the activation transition(s) of the receptor and the importance of this region in the determination of both GLP-1 peptide- and pathway-specific effects.

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Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences
Additional Information: © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
Uncontrolled Keywords: Biochemistry,Molecular Biology,Cell Biology
Publication ISSN: 1083-351X
Last Modified: 23 May 2024 07:14
Date Deposited: 05 Sep 2019 11:04
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
http://www.jbc. ... tent/287/6/3642 (Publisher URL)
PURE Output Type: Article
Published Date: 2012-02-03
Authors: Koole, Cassandra
Wootten, Denise
Simms, John (ORCID Profile 0000-0002-4675-0902)
Miller, Laurence J.
Christopoulos, Arthur
Sexton, Patrick M.



Version: Published Version

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