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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...

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Detalles Bibliográficos
Autores principales: de Hoog, Hans-Peter M., Lin JieRong, Esther M., Banerjee, Sourabh, Décaillot, Fabien M., Nallani, Madhavan
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
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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.
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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
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