Cargando…
Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes
G-protein coupled receptors (GPCRs) play a key role in physiological processes and are attractive drug targets. Their biophysical characterization is, however, highly challenging because of their innate instability outside a stabilizing membrane and the difficulty of finding a suitable expression sy...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4203850/ https://www.ncbi.nlm.nih.gov/pubmed/25329156 http://dx.doi.org/10.1371/journal.pone.0110847 |
_version_ | 1782340447362875392 |
---|---|
author | de Hoog, Hans-Peter M. Lin JieRong, Esther M. Banerjee, Sourabh Décaillot, Fabien M. Nallani, Madhavan |
author_facet | de Hoog, Hans-Peter M. Lin JieRong, Esther M. Banerjee, Sourabh Décaillot, Fabien M. Nallani, Madhavan |
author_sort | de Hoog, Hans-Peter M. |
collection | PubMed |
description | G-protein coupled receptors (GPCRs) play a key role in physiological processes and are attractive drug targets. Their biophysical characterization is, however, highly challenging because of their innate instability outside a stabilizing membrane and the difficulty of finding a suitable expression system. We here show the cell-free expression of a GPCR, CXCR4, and its direct embedding in diblock copolymer membranes. The polymer-stabilized CXCR4 is readily immobilized onto biosensor chips for label-free binding analysis. Kinetic characterization using a conformationally sensitive antibody shows the receptor to exist in the correctly folded conformation, showing binding behaviour that is commensurate with heterologously expressed CXCR4. |
format | Online Article Text |
id | pubmed-4203850 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42038502014-10-27 Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes de Hoog, Hans-Peter M. Lin JieRong, Esther M. Banerjee, Sourabh Décaillot, Fabien M. Nallani, Madhavan PLoS One Research Article G-protein coupled receptors (GPCRs) play a key role in physiological processes and are attractive drug targets. Their biophysical characterization is, however, highly challenging because of their innate instability outside a stabilizing membrane and the difficulty of finding a suitable expression system. We here show the cell-free expression of a GPCR, CXCR4, and its direct embedding in diblock copolymer membranes. The polymer-stabilized CXCR4 is readily immobilized onto biosensor chips for label-free binding analysis. Kinetic characterization using a conformationally sensitive antibody shows the receptor to exist in the correctly folded conformation, showing binding behaviour that is commensurate with heterologously expressed CXCR4. Public Library of Science 2014-10-20 /pmc/articles/PMC4203850/ /pubmed/25329156 http://dx.doi.org/10.1371/journal.pone.0110847 Text en © 2014 de Hoog 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 de Hoog, Hans-Peter M. Lin JieRong, Esther M. Banerjee, Sourabh Décaillot, Fabien M. Nallani, Madhavan Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title | Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title_full | Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title_fullStr | Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title_full_unstemmed | Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title_short | Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes |
title_sort | conformational antibody binding to a native, cell-free expressed gpcr in block copolymer membranes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4203850/ https://www.ncbi.nlm.nih.gov/pubmed/25329156 http://dx.doi.org/10.1371/journal.pone.0110847 |
work_keys_str_mv | AT dehooghanspeterm conformationalantibodybindingtoanativecellfreeexpressedgpcrinblockcopolymermembranes AT linjierongestherm conformationalantibodybindingtoanativecellfreeexpressedgpcrinblockcopolymermembranes AT banerjeesourabh conformationalantibodybindingtoanativecellfreeexpressedgpcrinblockcopolymermembranes AT decaillotfabienm conformationalantibodybindingtoanativecellfreeexpressedgpcrinblockcopolymermembranes AT nallanimadhavan conformationalantibodybindingtoanativecellfreeexpressedgpcrinblockcopolymermembranes |