Document Detail

Quantification of ligand binding to G-protein coupled receptors on cell membranes by ellipsometry.
MedLine Citation:
PMID:  23049983     Owner:  NLM     Status:  MEDLINE    
G-protein-coupled receptors (GPCRs) are prime drug targets and targeted by approximately 60% of current therapeutic drugs such as β-blockers, antipsychotics and analgesics. However, no biophysical methods are available to quantify their interactions with ligand binding in a native environment. Here, we use ellipsometry to quantify specific interactions of receptors within native cell membranes. As a model system, the GPCR-ligand CXCL12α and its receptor CXCR4 are used. Human-derived Ishikawa cells were deposited onto gold coated slides via Langmuir-Schaefer film deposition and interactions between the receptor CXCR4 on these cells and its ligand CXCL12α were detected via total internal reflection ellipsometry (TIRE). This interaction could be inhibited by application of the CXCR4-binding drug AMD3100. Advantages of this approach are that it allows measurement of interactions in a lipid environment without the need for labelling, protein purification or reconstitution of membrane proteins. This technique is potentially applicable to a wide variety of cell types and their membrane receptors, providing a novel method to determine ligand or drug interactions targeting GPCRs and other membrane proteins.
Verena Kriechbaumer; Alexei Nabok; Robert Widdowson; David P Smith; Ben M Abell
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-09-26
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-10-10     Completed Date:  2013-04-10     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e46221     Citation Subset:  IM    
Biomedical Research Centre, Sheffield Hallam University, Sheffield, United Kingdom.
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MeSH Terms
Cell Line
Cell Membrane / genetics,  metabolism*
Chemokine CXCL12 / genetics,  metabolism
Heterocyclic Compounds / pharmacology
Microscopy, Confocal
Protein Binding / drug effects
Receptors, CXCR / genetics,  metabolism*
Receptors, CXCR4 / genetics,  metabolism*
Receptors, G-Protein-Coupled / genetics,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Grant Support
BB/E01559X/1//Biotechnology and Biological Sciences Research Council
Reg. No./Substance:
0/CXCL12 protein, human; 0/CXCR4 protein, human; 0/CXCR7 protein, human; 0/Chemokine CXCL12; 0/Heterocyclic Compounds; 0/Receptors, CXCR; 0/Receptors, CXCR4; 0/Receptors, G-Protein-Coupled; 155148-31-5/JM 3100

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