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Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP
It is well-established that activation of heterotrimeric G-proteins (Gαβγ) by G-protein-coupled receptors (GPCRs) stimulated by neurotransmitters is a key mechanism underlying neuromodulation. Much less is known about how G-protein regulation after receptor-mediated activation contributes to neuromo...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10153262/ https://www.ncbi.nlm.nih.gov/pubmed/37131787 http://dx.doi.org/10.1101/2023.04.20.537566 |
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author | Luebbers, Alex Zhou, Myles Eyles, Stephen J Garcia-Marcos, Mikel |
author_facet | Luebbers, Alex Zhou, Myles Eyles, Stephen J Garcia-Marcos, Mikel |
author_sort | Luebbers, Alex |
collection | PubMed |
description | It is well-established that activation of heterotrimeric G-proteins (Gαβγ) by G-protein-coupled receptors (GPCRs) stimulated by neurotransmitters is a key mechanism underlying neuromodulation. Much less is known about how G-protein regulation after receptor-mediated activation contributes to neuromodulation. Recent evidence indicates that the neuronal protein GINIP shapes GPCR inhibitory neuromodulation via a unique mechanism of G-protein regulation that controls neurological processes like pain and seizure susceptibility. However, the molecular basis of this mechanism remains ill-defined because the structural determinants of GINIP responsible for binding Gαi subunits and regulating G-protein signaling are not known. Here, we combined hydrogen-deuterium exchange mass-spectrometry, protein folding predictions, bioluminescence resonance energy transfer assays, and biochemical experiments to identify the first loop of the PHD domain of GINIP as an obligatory requirement for Gαi binding. Surprisingly, our results support a model in which GINIP undergoes a long-range conformational change to accommodate Gαi binding to this loop. Using cell-based assays, we demonstrate that specific amino acids in the first loop of the PHD domain are essential for the regulation of Gαi-GTP and free Gβγ signaling upon neurotransmitter GPCR stimulation. In summary, these findings shed light onto the molecular basis for a post-receptor mechanism of G-protein regulation that fine-tunes inhibitory neuromodulation. |
format | Online Article Text |
id | pubmed-10153262 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-101532622023-05-03 Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP Luebbers, Alex Zhou, Myles Eyles, Stephen J Garcia-Marcos, Mikel bioRxiv Article It is well-established that activation of heterotrimeric G-proteins (Gαβγ) by G-protein-coupled receptors (GPCRs) stimulated by neurotransmitters is a key mechanism underlying neuromodulation. Much less is known about how G-protein regulation after receptor-mediated activation contributes to neuromodulation. Recent evidence indicates that the neuronal protein GINIP shapes GPCR inhibitory neuromodulation via a unique mechanism of G-protein regulation that controls neurological processes like pain and seizure susceptibility. However, the molecular basis of this mechanism remains ill-defined because the structural determinants of GINIP responsible for binding Gαi subunits and regulating G-protein signaling are not known. Here, we combined hydrogen-deuterium exchange mass-spectrometry, protein folding predictions, bioluminescence resonance energy transfer assays, and biochemical experiments to identify the first loop of the PHD domain of GINIP as an obligatory requirement for Gαi binding. Surprisingly, our results support a model in which GINIP undergoes a long-range conformational change to accommodate Gαi binding to this loop. Using cell-based assays, we demonstrate that specific amino acids in the first loop of the PHD domain are essential for the regulation of Gαi-GTP and free Gβγ signaling upon neurotransmitter GPCR stimulation. In summary, these findings shed light onto the molecular basis for a post-receptor mechanism of G-protein regulation that fine-tunes inhibitory neuromodulation. Cold Spring Harbor Laboratory 2023-04-21 /pmc/articles/PMC10153262/ /pubmed/37131787 http://dx.doi.org/10.1101/2023.04.20.537566 Text en https://creativecommons.org/licenses/by-nc/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator. |
spellingShingle | Article Luebbers, Alex Zhou, Myles Eyles, Stephen J Garcia-Marcos, Mikel Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title | Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title_full | Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title_fullStr | Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title_full_unstemmed | Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title_short | Dissecting the molecular basis for the modulation of neurotransmitter GPCR signaling by GINIP |
title_sort | dissecting the molecular basis for the modulation of neurotransmitter gpcr signaling by ginip |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10153262/ https://www.ncbi.nlm.nih.gov/pubmed/37131787 http://dx.doi.org/10.1101/2023.04.20.537566 |
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