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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 Shukla, Arun

PloS one, 2017-11, Vol.12 (11), p.e0187306-e0187306 [Periódico revisado por pares]

United States: Public Library of Science

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Título:
Generation of G protein-coupled receptor antibodies differentially sensitive to conformational states
Autor: Heimann, Andrea S ; Gupta, Achla ; Gomes, Ivone ; Rayees, Rahman ; Schlessinger, Avner ; Ferro, Emer S ; Unterwald, Ellen M ; Devi, Lakshmi A
Shukla, Arun
Assuntos: Animals ; Antibodies ; Antibodies - immunology ; Biology and Life Sciences ; Brain ; Change detection ; Cocaine ; Dopamine ; Dopamine receptors ; Dopamine transporter ; Dosage and administration ; Enzyme-Linked Immunosorbent Assay ; Epitopes ; Epitopes - chemistry ; Epitopes - immunology ; G protein-coupled receptors ; G proteins ; Glutamate ; HEK293 Cells ; Humans ; Immunoglobulins ; Male ; Medicine ; Medicine and Health Sciences ; Narcotics ; Peptides ; Physical Sciences ; Protein Conformation ; Proteins ; Rabbits ; Rats ; Rats, Wistar ; Receptors ; Receptors, G-Protein-Coupled - agonists ; Receptors, G-Protein-Coupled - antagonists & inhibitors ; Receptors, G-Protein-Coupled - chemistry ; Receptors, G-Protein-Coupled - immunology ; Recognition ; Research and Analysis Methods ; Signal transduction ; Studies ; Viral antibodies
É parte de: PloS one, 2017-11, Vol.12 (11), p.e0187306-e0187306
Notas: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
Competing Interests: The authors have declared that no competing interests exist.
Descrição: 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.
Editor: United States: Public Library of Science
Idioma: Inglês

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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 Shukla, Arun

United States: Public Library of Science

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