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Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction

The apelin and Elabela proteins constitute a spatiotemporal double-ligand system that controls apelin receptor (APJ) signal transduction. Phosphorylation of multiple sites within the C-terminus of APJ is essential for the recruitment of β-arrestins. We sought to determine the precise mechanisms by w...

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Autores principales: Chen, Jing, Chen, Xiaoyu, Li, Sheng, Jiang, Yunlu, Mao, Huiling, Zhang, Rumin, Ji, Bingyuan, Yan, Maocai, Cai, Xin, Wang, Chunmei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338617/
https://www.ncbi.nlm.nih.gov/pubmed/32863206
http://dx.doi.org/10.1016/j.redox.2020.101629
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author Chen, Jing
Chen, Xiaoyu
Li, Sheng
Jiang, Yunlu
Mao, Huiling
Zhang, Rumin
Ji, Bingyuan
Yan, Maocai
Cai, Xin
Wang, Chunmei
author_facet Chen, Jing
Chen, Xiaoyu
Li, Sheng
Jiang, Yunlu
Mao, Huiling
Zhang, Rumin
Ji, Bingyuan
Yan, Maocai
Cai, Xin
Wang, Chunmei
author_sort Chen, Jing
collection PubMed
description The apelin and Elabela proteins constitute a spatiotemporal double-ligand system that controls apelin receptor (APJ) signal transduction. Phosphorylation of multiple sites within the C-terminus of APJ is essential for the recruitment of β-arrestins. We sought to determine the precise mechanisms by which apelin and Elabela promote APJ phosphorylation, and to elucidate the influence of β-arrestin phosphorylation on G-protein-coupled receptor (GPCR)/β-arrestin-dependent signaling. We used techniques including mass spectrometry (MS), mutation analysis, and bioluminescence resonance energy transfer (BRET) to evaluate the role of phosphorylation sites in APJ-mediated G-protein-dependent and β-dependent signaling. Phosphorylation of APJ occurred at five serine residues in the C-terminal region (Ser335, Ser339, Ser345, Ser348 and Ser369). We also identified two phosphorylation sites in β-arrestin1 and three in β-arrestin2, including three previously identified residues (Ser412, Ser361, and Thr383) and two new sites, Tyr47 in β-arrestin1 and Tyr48 in β-arrestin2. APJ mutations did not affect the phosphorylation of β-arrestins, but it affects the β-arrestin signaling pathway, specifically Ser335 and Ser339. Mutation of Ser335 decreased the ability of the receptor to interact with β-arrestin1/2 and AP2, indicating that APJ affects the β-arrestin signaling pathway by stimulating Elabela. Mutation of Ser339 abolished the capability of the receptor to interact with GRK2 and β-arrestin1/2 upon stimulation with apelin-36, and disrupted receptor internalization and β-arrestin-dependent ERK1/2 activation. Five peptides act on distinct phosphorylation sites at the APJ C-terminus, differentially regulating APJ signal transduction and causing different biological effects. These findings may facilitate screening for drugs to treat cardiovascular and metabolic diseases.
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spelling pubmed-73386172020-07-14 Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction Chen, Jing Chen, Xiaoyu Li, Sheng Jiang, Yunlu Mao, Huiling Zhang, Rumin Ji, Bingyuan Yan, Maocai Cai, Xin Wang, Chunmei Redox Biol Research Paper The apelin and Elabela proteins constitute a spatiotemporal double-ligand system that controls apelin receptor (APJ) signal transduction. Phosphorylation of multiple sites within the C-terminus of APJ is essential for the recruitment of β-arrestins. We sought to determine the precise mechanisms by which apelin and Elabela promote APJ phosphorylation, and to elucidate the influence of β-arrestin phosphorylation on G-protein-coupled receptor (GPCR)/β-arrestin-dependent signaling. We used techniques including mass spectrometry (MS), mutation analysis, and bioluminescence resonance energy transfer (BRET) to evaluate the role of phosphorylation sites in APJ-mediated G-protein-dependent and β-dependent signaling. Phosphorylation of APJ occurred at five serine residues in the C-terminal region (Ser335, Ser339, Ser345, Ser348 and Ser369). We also identified two phosphorylation sites in β-arrestin1 and three in β-arrestin2, including three previously identified residues (Ser412, Ser361, and Thr383) and two new sites, Tyr47 in β-arrestin1 and Tyr48 in β-arrestin2. APJ mutations did not affect the phosphorylation of β-arrestins, but it affects the β-arrestin signaling pathway, specifically Ser335 and Ser339. Mutation of Ser335 decreased the ability of the receptor to interact with β-arrestin1/2 and AP2, indicating that APJ affects the β-arrestin signaling pathway by stimulating Elabela. Mutation of Ser339 abolished the capability of the receptor to interact with GRK2 and β-arrestin1/2 upon stimulation with apelin-36, and disrupted receptor internalization and β-arrestin-dependent ERK1/2 activation. Five peptides act on distinct phosphorylation sites at the APJ C-terminus, differentially regulating APJ signal transduction and causing different biological effects. These findings may facilitate screening for drugs to treat cardiovascular and metabolic diseases. Elsevier 2020-06-30 /pmc/articles/PMC7338617/ /pubmed/32863206 http://dx.doi.org/10.1016/j.redox.2020.101629 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Paper
Chen, Jing
Chen, Xiaoyu
Li, Sheng
Jiang, Yunlu
Mao, Huiling
Zhang, Rumin
Ji, Bingyuan
Yan, Maocai
Cai, Xin
Wang, Chunmei
Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title_full Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title_fullStr Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title_full_unstemmed Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title_short Individual phosphorylation sites at the C-terminus of the apelin receptor play different roles in signal transduction
title_sort individual phosphorylation sites at the c-terminus of the apelin receptor play different roles in signal transduction
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338617/
https://www.ncbi.nlm.nih.gov/pubmed/32863206
http://dx.doi.org/10.1016/j.redox.2020.101629
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