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Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states

The N-terminal region of G protein-coupled receptors can be efficiently targeted for the generation of receptor-selective antibodies. These antibodies are useful for the biochemical characterization of the receptors. In this study, we developed a set of criteria to select the optimal epitope and app...

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Autores principales: Heimann, Andrea S., Gupta, Achla, Gomes, Ivone, Rayees, Rahman, Schlessinger, Avner, Ferro, Emer S., Unterwald, Ellen M., Devi, Lakshmi A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5665533/
https://www.ncbi.nlm.nih.gov/pubmed/29091950
http://dx.doi.org/10.1371/journal.pone.0187306
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author Heimann, Andrea S.
Gupta, Achla
Gomes, Ivone
Rayees, Rahman
Schlessinger, Avner
Ferro, Emer S.
Unterwald, Ellen M.
Devi, Lakshmi A.
author_facet Heimann, Andrea S.
Gupta, Achla
Gomes, Ivone
Rayees, Rahman
Schlessinger, Avner
Ferro, Emer S.
Unterwald, Ellen M.
Devi, Lakshmi A.
author_sort Heimann, Andrea S.
collection PubMed
description The N-terminal region of G protein-coupled receptors can be efficiently targeted for the generation of receptor-selective antibodies. These antibodies are useful for the biochemical characterization of the receptors. In this study, we developed a set of criteria to select the optimal epitope and applied them to generate antibodies to the N-terminal region of 34 different G protein-coupled receptors. The antibody characterization revealed that a subset of antibodies exhibited increased recognition of the receptor following agonist treatment and this increase could be blocked by treatment with the receptor antagonist. An analysis of the epitopes showed that those antibodies that exhibit increased recognition are on average twelve residues long, have an overall net negative charge and are enriched in aspartic and glutamic acids. These antibodies are useful since they facilitate studies examining dose dependent increases in recognition of receptors in heterologous cells as well as in native tissue. Another interesting use of these antibodies is that they facilitate measuring changes in receptor recognition in brain following peripheral drug administration; for example, systemic administration of cocaine, a blocker of dopamine transporter that increases local dopamine levels at the synapse, was found to lead to increases in antibody recognition of dopamine receptors in the brain. Taken together these studies, in addition to describing novel tools to study native receptors, provide a framework for the generation of antibodies to G protein-coupled receptors that can detect ligand-induced conformational changes.
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spelling pubmed-56655332017-11-09 Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states Heimann, Andrea S. Gupta, Achla Gomes, Ivone Rayees, Rahman Schlessinger, Avner Ferro, Emer S. Unterwald, Ellen M. Devi, Lakshmi A. PLoS One Research Article The N-terminal region of G protein-coupled receptors can be efficiently targeted for the generation of receptor-selective antibodies. These antibodies are useful for the biochemical characterization of the receptors. In this study, we developed a set of criteria to select the optimal epitope and applied them to generate antibodies to the N-terminal region of 34 different G protein-coupled receptors. The antibody characterization revealed that a subset of antibodies exhibited increased recognition of the receptor following agonist treatment and this increase could be blocked by treatment with the receptor antagonist. An analysis of the epitopes showed that those antibodies that exhibit increased recognition are on average twelve residues long, have an overall net negative charge and are enriched in aspartic and glutamic acids. These antibodies are useful since they facilitate studies examining dose dependent increases in recognition of receptors in heterologous cells as well as in native tissue. Another interesting use of these antibodies is that they facilitate measuring changes in receptor recognition in brain following peripheral drug administration; for example, systemic administration of cocaine, a blocker of dopamine transporter that increases local dopamine levels at the synapse, was found to lead to increases in antibody recognition of dopamine receptors in the brain. Taken together these studies, in addition to describing novel tools to study native receptors, provide a framework for the generation of antibodies to G protein-coupled receptors that can detect ligand-induced conformational changes. Public Library of Science 2017-11-01 /pmc/articles/PMC5665533/ /pubmed/29091950 http://dx.doi.org/10.1371/journal.pone.0187306 Text en © 2017 Heimann 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Heimann, Andrea S.
Gupta, Achla
Gomes, Ivone
Rayees, Rahman
Schlessinger, Avner
Ferro, Emer S.
Unterwald, Ellen M.
Devi, Lakshmi A.
Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title_full Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title_fullStr Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title_full_unstemmed Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title_short Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
title_sort generation of g protein-coupled receptor antibodies differentially sensitive to conformational states
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5665533/
https://www.ncbi.nlm.nih.gov/pubmed/29091950
http://dx.doi.org/10.1371/journal.pone.0187306
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