Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry
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,...
| Main Authors: | , , , , |
|---|---|
| Format: | Online |
| Language: | English |
| Published: |
Public Library of Science
2012
|
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3458955/ |
| id |
pubmed-3458955 |
|---|---|
| recordtype |
oai_dc |
| spelling |
pubmed-34589552012-10-03 Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry Kriechbaumer, Verena Nabok, Alexei Widdowson, Robert Smith, David P. Abell, Ben M. Research Article 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. Public Library of Science 2012-09-26 /pmc/articles/PMC3458955/ /pubmed/23049983 http://dx.doi.org/10.1371/journal.pone.0046221 Text en © 2012 Kriechbaumer 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. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Kriechbaumer, Verena Nabok, Alexei Widdowson, Robert Smith, David P. Abell, Ben M. |
| spellingShingle |
Kriechbaumer, Verena Nabok, Alexei Widdowson, Robert Smith, David P. Abell, Ben M. Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| author_facet |
Kriechbaumer, Verena Nabok, Alexei Widdowson, Robert Smith, David P. Abell, Ben M. |
| author_sort |
Kriechbaumer, Verena |
| title |
Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| title_short |
Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| title_full |
Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| title_fullStr |
Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| title_full_unstemmed |
Quantification of Ligand Binding to G-Protein Coupled Receptors on Cell Membranes by Ellipsometry |
| title_sort |
quantification of ligand binding to g-protein coupled receptors on cell membranes by ellipsometry |
| description |
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. |
| publisher |
Public Library of Science |
| publishDate |
2012 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3458955/ |
| _version_ |
1611912123933261824 |