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Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes
Posted: October 20, 2014
PLoS ONE 9(10): e110847 © 2014 de Hoog et al
doi: https://doi.org/10.1371/journal.pone.0110847
Authors: Hans-Peter M. de Hoog,Esther M. Lin JieRong,Sourabh Banerjee,Fabien M. Décaillot,Madhavan Nallani
Abstract
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.