Cargando…
Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids
G-protein coupled receptors (GPCRs) are important therapeutic targets since more than 40% of the drugs on the market exert their action through these proteins. To decipher the molecular mechanisms of activation and signaling, GPCRs often need to be isolated and reconstituted from a detergent-solubil...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5099921/ https://www.ncbi.nlm.nih.gov/pubmed/27824122 http://dx.doi.org/10.1038/srep36181 |
_version_ | 1782466029650182144 |
---|---|
author | Calmet, Pierre De Maria, Monica Harté, Etienne Lamb, Daniel Serrano-Vega, Maria Jazayeri, Ali Tschammer, Nuska Alves, Isabel D. |
author_facet | Calmet, Pierre De Maria, Monica Harté, Etienne Lamb, Daniel Serrano-Vega, Maria Jazayeri, Ali Tschammer, Nuska Alves, Isabel D. |
author_sort | Calmet, Pierre |
collection | PubMed |
description | G-protein coupled receptors (GPCRs) are important therapeutic targets since more than 40% of the drugs on the market exert their action through these proteins. To decipher the molecular mechanisms of activation and signaling, GPCRs often need to be isolated and reconstituted from a detergent-solubilized state into a well-defined and controllable lipid model system. Several methods exist to reconstitute membrane proteins in lipid systems but usually the reconstitution success is tested at the end of the experiment and often by an additional and indirect method. Irrespective of the method used, the reconstitution process is often an intractable and time-consuming trial-and-error procedure. Herein, we present a method that allows directly monitoring the reconstitution of GPCRs in model planar lipid membranes. Plasmon waveguide resonance (PWR) allows following GPCR lipid reconstitution process without any labeling and with high sensitivity. Additionally, the method is ideal to probe the lipid effect on receptor ligand binding as demonstrated by antagonist binding to the chemokine CCR5 receptor. |
format | Online Article Text |
id | pubmed-5099921 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-50999212016-11-14 Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids Calmet, Pierre De Maria, Monica Harté, Etienne Lamb, Daniel Serrano-Vega, Maria Jazayeri, Ali Tschammer, Nuska Alves, Isabel D. Sci Rep Article G-protein coupled receptors (GPCRs) are important therapeutic targets since more than 40% of the drugs on the market exert their action through these proteins. To decipher the molecular mechanisms of activation and signaling, GPCRs often need to be isolated and reconstituted from a detergent-solubilized state into a well-defined and controllable lipid model system. Several methods exist to reconstitute membrane proteins in lipid systems but usually the reconstitution success is tested at the end of the experiment and often by an additional and indirect method. Irrespective of the method used, the reconstitution process is often an intractable and time-consuming trial-and-error procedure. Herein, we present a method that allows directly monitoring the reconstitution of GPCRs in model planar lipid membranes. Plasmon waveguide resonance (PWR) allows following GPCR lipid reconstitution process without any labeling and with high sensitivity. Additionally, the method is ideal to probe the lipid effect on receptor ligand binding as demonstrated by antagonist binding to the chemokine CCR5 receptor. Nature Publishing Group 2016-11-08 /pmc/articles/PMC5099921/ /pubmed/27824122 http://dx.doi.org/10.1038/srep36181 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Calmet, Pierre De Maria, Monica Harté, Etienne Lamb, Daniel Serrano-Vega, Maria Jazayeri, Ali Tschammer, Nuska Alves, Isabel D. Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title | Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title_full | Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title_fullStr | Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title_full_unstemmed | Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title_short | Real time monitoring of membrane GPCR reconstitution by plasmon waveguide resonance: on the role of lipids |
title_sort | real time monitoring of membrane gpcr reconstitution by plasmon waveguide resonance: on the role of lipids |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5099921/ https://www.ncbi.nlm.nih.gov/pubmed/27824122 http://dx.doi.org/10.1038/srep36181 |
work_keys_str_mv | AT calmetpierre realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT demariamonica realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT harteetienne realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT lambdaniel realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT serranovegamaria realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT jazayeriali realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT tschammernuska realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids AT alvesisabeld realtimemonitoringofmembranegpcrreconstitutionbyplasmonwaveguideresonanceontheroleoflipids |