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Conformational antibody binding to a native, cell-free expressed GPCR in block copolymer membranes

Authors :
Hans-Peter M. de Hoog
Sourabh Banerjee
Fabien Decaillot
Esther M. Lin JieRong
Madhavan Nallani
Karnik, Sadashiva
School of Materials Science & Engineering
Source :
PLoS ONE, Vol 9, Iss 10, p e110847 (2014), PLoS ONE
Publication Year :
2014
Publisher :
Public Library of Science (PLoS), 2014.

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. ASTAR (Agency for Sci., Tech. and Research, S’pore) Published version

Details

Language :
English
ISSN :
19326203
Volume :
9
Issue :
10
Database :
OpenAIRE
Journal :
PLoS ONE
Accession number :
edsair.doi.dedup.....c6ada90cebd3c8c7b2546af8c7cd340c