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Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway

Aiysha Thompson, Venkat Kanamarlapudi Orcid Logo

Biochemical Pharmacology, Volume: 93, Issue: 1, Pages: 72 - 84

Swansea University Author: Venkat Kanamarlapudi Orcid Logo

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DOI (Published version): 10.1016/j.bcp.2014.10.015

Abstract

The glucagon like peptide-1 receptor (GLP-1R) is a G-protein coupled receptor (GPCR) and an important target in the treatment of type 2 diabetes mellitus (T2DM). Upon stimulation with agonist, the GLP-1R signals through both Gαs and Gαq coupled pathways to stimulate insulin secretion. The agonist in...

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Published in: Biochemical Pharmacology
Published: 2015
URI: https://cronfa.swan.ac.uk/Record/cronfa19150
first_indexed 2015-01-22T02:56:11Z
last_indexed 2019-06-21T19:30:23Z
id cronfa19150
recordtype SURis
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spelling 2019-06-21T15:14:01.6361580 v2 19150 2014-11-10 Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway 63741801137148abfa4c00cd547dcdfa 0000-0002-8739-1483 Venkat Kanamarlapudi Venkat Kanamarlapudi true false 2014-11-10 MEDS The glucagon like peptide-1 receptor (GLP-1R) is a G-protein coupled receptor (GPCR) and an important target in the treatment of type 2 diabetes mellitus (T2DM). Upon stimulation with agonist, the GLP-1R signals through both Gαs and Gαq coupled pathways to stimulate insulin secretion. The agonist induced GLP-1R internalisation has recently been shown to be important for insulin secretion. However, the molecular mechanisms underlying GLP-1R internalisation remain unknown. The aim of this study was to determine the role of GLP-1R downstream signalling pathways in its internalisation. Agonist induced human GLP-1R (hGLP-1R) internalisation and activity were examined using a number of techniques including immunoblotting, ELISA, immunofluorescence, and luciferase assays to determine cAMP production, intracellular Ca2+ accumulation and ERK phosphorylation. Agonist induced hGLP-1R internalisation is dependent on caveolin-1 and dynamin. Inhibition of the Gαq pathway but not the Gαs pathway affected hGLP-1R internalisation. Consistent with this, hGLP-1R mutant T149 M and small molecule agonists (compound 2 and compound B), which activate only the Gαs pathway, failed to induce internalisation of the receptor. Chemical inhibitors of the Gαq pathway, PKC and ERK phosphorylation significantly reduced agonist induced hGLP-1R internalisation. These inhibitors also suppressed agonist induced ERK1/2 phosphorylation demonstrating that the phosphorylated ERK acts downstream of the Gαq pathway in the hGLP-1R internalisation. In summary, agonist induced hGLP-1R internalisation is mediated by the Gαq pathway. The internalised hGLP-1R stimulates insulin secretion from pancreatic β-cells, indicating the importance of GLP-1 internalisation for insulin secretion. Journal Article Biochemical Pharmacology 93 1 72 84 Glucagon like peptide-1 (GLP-1); GLP-1 receptor (GLP-1R); G protein coupled receptor (GPCR); Endocytosis; Gαq pathway 1 1 2015 2015-01-01 10.1016/j.bcp.2014.10.015 COLLEGE NANME Medical School COLLEGE CODE MEDS Swansea University 2019-06-21T15:14:01.6361580 2014-11-10T09:50:19.6985418 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Aiysha Thompson 1 Venkat Kanamarlapudi 0000-0002-8739-1483 2
title Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
spellingShingle Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
Venkat Kanamarlapudi
title_short Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
title_full Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
title_fullStr Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
title_full_unstemmed Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
title_sort Agonist-induced internalisation of the glucagon-like peptide-1 receptor is mediated by the Gαq pathway
author_id_str_mv 63741801137148abfa4c00cd547dcdfa
author_id_fullname_str_mv 63741801137148abfa4c00cd547dcdfa_***_Venkat Kanamarlapudi
author Venkat Kanamarlapudi
author2 Aiysha Thompson
Venkat Kanamarlapudi
format Journal article
container_title Biochemical Pharmacology
container_volume 93
container_issue 1
container_start_page 72
publishDate 2015
institution Swansea University
doi_str_mv 10.1016/j.bcp.2014.10.015
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
active_str 0
description The glucagon like peptide-1 receptor (GLP-1R) is a G-protein coupled receptor (GPCR) and an important target in the treatment of type 2 diabetes mellitus (T2DM). Upon stimulation with agonist, the GLP-1R signals through both Gαs and Gαq coupled pathways to stimulate insulin secretion. The agonist induced GLP-1R internalisation has recently been shown to be important for insulin secretion. However, the molecular mechanisms underlying GLP-1R internalisation remain unknown. The aim of this study was to determine the role of GLP-1R downstream signalling pathways in its internalisation. Agonist induced human GLP-1R (hGLP-1R) internalisation and activity were examined using a number of techniques including immunoblotting, ELISA, immunofluorescence, and luciferase assays to determine cAMP production, intracellular Ca2+ accumulation and ERK phosphorylation. Agonist induced hGLP-1R internalisation is dependent on caveolin-1 and dynamin. Inhibition of the Gαq pathway but not the Gαs pathway affected hGLP-1R internalisation. Consistent with this, hGLP-1R mutant T149 M and small molecule agonists (compound 2 and compound B), which activate only the Gαs pathway, failed to induce internalisation of the receptor. Chemical inhibitors of the Gαq pathway, PKC and ERK phosphorylation significantly reduced agonist induced hGLP-1R internalisation. These inhibitors also suppressed agonist induced ERK1/2 phosphorylation demonstrating that the phosphorylated ERK acts downstream of the Gαq pathway in the hGLP-1R internalisation. In summary, agonist induced hGLP-1R internalisation is mediated by the Gαq pathway. The internalised hGLP-1R stimulates insulin secretion from pancreatic β-cells, indicating the importance of GLP-1 internalisation for insulin secretion.
published_date 2015-01-01T06:38:59Z
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