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Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor

G protein–coupled receptors (GPCRs) signal through a limited number of G-protein pathways and play crucial roles in many biological processes. Studies of their in vivo functions have been hampered by the molecular and functional diversity of GPCRs and the paucity of ligands with specific signaling e...

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Autores principales: Chang, Wei Chun, Ng, Jennifer K., Nguyen, Trieu, Pellissier, Lucie, Claeysen, Sylvie, Hsiao, Edward C., Conklin, Bruce R.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2007
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267039/
https://www.ncbi.nlm.nih.gov/pubmed/18338032
http://dx.doi.org/10.1371/journal.pone.0001317
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author Chang, Wei Chun
Ng, Jennifer K.
Nguyen, Trieu
Pellissier, Lucie
Claeysen, Sylvie
Hsiao, Edward C.
Conklin, Bruce R.
author_facet Chang, Wei Chun
Ng, Jennifer K.
Nguyen, Trieu
Pellissier, Lucie
Claeysen, Sylvie
Hsiao, Edward C.
Conklin, Bruce R.
author_sort Chang, Wei Chun
collection PubMed
description G protein–coupled receptors (GPCRs) signal through a limited number of G-protein pathways and play crucial roles in many biological processes. Studies of their in vivo functions have been hampered by the molecular and functional diversity of GPCRs and the paucity of ligands with specific signaling effects. To better compare the effects of activating different G-protein signaling pathways through ligand-induced or constitutive signaling, we developed a new series of RASSLs (receptors activated solely by synthetic ligands) that activate different G-protein signaling pathways. These RASSLs are based on the human 5-HT(4b) receptor, a GPCR with high constitutive G(s) signaling and strong ligand-induced G-protein activation of the G(s) and G(s/q) pathways. The first receptor in this series, 5-HT(4)-D(100)A or Rs1 (RASSL serotonin 1), is not activated by its endogenous agonist, serotonin, but is selectively activated by the small synthetic molecules GR113808, GR125487, and RO110-0235. All agonists potently induced G(s) signaling, but only a few (e.g., zacopride) also induced signaling via the G(q) pathway. Zacopride-induced G(q) signaling was enhanced by replacing the C-terminus of Rs1 with the C-terminus of the human 5-HT(2C) receptor. Additional point mutations (D(66)A and D(66)N) blocked constitutive G(s) signaling and lowered ligand-induced G(q) signaling. Replacing the third intracellular loop of Rs1 with that of human 5-HT(1A) conferred ligand-mediated G(i) signaling. This G(i)-coupled RASSL, Rs1.3, exhibited no measurable signaling to the G(s) or G(q) pathway. These findings show that the signaling repertoire of Rs1 can be expanded and controlled by receptor engineering and drug selection.
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spelling pubmed-22670392008-03-12 Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor Chang, Wei Chun Ng, Jennifer K. Nguyen, Trieu Pellissier, Lucie Claeysen, Sylvie Hsiao, Edward C. Conklin, Bruce R. PLoS One Research Article G protein–coupled receptors (GPCRs) signal through a limited number of G-protein pathways and play crucial roles in many biological processes. Studies of their in vivo functions have been hampered by the molecular and functional diversity of GPCRs and the paucity of ligands with specific signaling effects. To better compare the effects of activating different G-protein signaling pathways through ligand-induced or constitutive signaling, we developed a new series of RASSLs (receptors activated solely by synthetic ligands) that activate different G-protein signaling pathways. These RASSLs are based on the human 5-HT(4b) receptor, a GPCR with high constitutive G(s) signaling and strong ligand-induced G-protein activation of the G(s) and G(s/q) pathways. The first receptor in this series, 5-HT(4)-D(100)A or Rs1 (RASSL serotonin 1), is not activated by its endogenous agonist, serotonin, but is selectively activated by the small synthetic molecules GR113808, GR125487, and RO110-0235. All agonists potently induced G(s) signaling, but only a few (e.g., zacopride) also induced signaling via the G(q) pathway. Zacopride-induced G(q) signaling was enhanced by replacing the C-terminus of Rs1 with the C-terminus of the human 5-HT(2C) receptor. Additional point mutations (D(66)A and D(66)N) blocked constitutive G(s) signaling and lowered ligand-induced G(q) signaling. Replacing the third intracellular loop of Rs1 with that of human 5-HT(1A) conferred ligand-mediated G(i) signaling. This G(i)-coupled RASSL, Rs1.3, exhibited no measurable signaling to the G(s) or G(q) pathway. These findings show that the signaling repertoire of Rs1 can be expanded and controlled by receptor engineering and drug selection. Public Library of Science 2007-12-19 /pmc/articles/PMC2267039/ /pubmed/18338032 http://dx.doi.org/10.1371/journal.pone.0001317 Text en Chang et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chang, Wei Chun
Ng, Jennifer K.
Nguyen, Trieu
Pellissier, Lucie
Claeysen, Sylvie
Hsiao, Edward C.
Conklin, Bruce R.
Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title_full Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title_fullStr Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title_full_unstemmed Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title_short Modifying Ligand-Induced and Constitutive Signaling of the Human 5-HT(4) Receptor
title_sort modifying ligand-induced and constitutive signaling of the human 5-ht(4) receptor
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2267039/
https://www.ncbi.nlm.nih.gov/pubmed/18338032
http://dx.doi.org/10.1371/journal.pone.0001317
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