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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
Descripción
Sumario: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.