Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue

The forward (kon) and reverse (koff) rate constants of drug-target interactions have important implications for therapeutic efficacy. Hence, time-resolved assays capable of measuring these binding rate constants may be informative to drug discovery efforts. Here, we used an ion channel activation as...

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Autores: Ågren, Richard, Stepniewski, Tomasz Maciej, 1988-, Zeberg, Hugo, Selent, Jana, Sahlholm, Kristoffer
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/47347
Acesso em linha:http://hdl.handle.net/10230/47347
http://dx.doi.org/10.3390/ijms22084078
Access Level:acceso abierto
Palavra-chave:G protein-coupled receptor
Xenopus oocytes
Aminotetralins
Electrophysiology
Molecular dynamics simulation
Phenethylamines
Voltage-clamp
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oai_identifier_str oai:recercat.cat:10230/47347
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spelling Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residueÅgren, RichardStepniewski, Tomasz Maciej, 1988-Zeberg, HugoSelent, JanaSahlholm, KristofferG protein-coupled receptorXenopus oocytesAminotetralinsElectrophysiologyMolecular dynamics simulationPhenethylaminesVoltage-clampThe forward (kon) and reverse (koff) rate constants of drug-target interactions have important implications for therapeutic efficacy. Hence, time-resolved assays capable of measuring these binding rate constants may be informative to drug discovery efforts. Here, we used an ion channel activation assay to estimate the kons and koffs of four dopamine D2 receptor (D2R) agonists; dopamine (DA), p-tyramine, (R)- and (S)-5-OH-dipropylaminotetralin (DPAT). We further probed the role of the conserved serine S1935.42 by mutagenesis, taking advantage of the preferential interaction of (S)-, but not (R)-5-OH-DPAT with this residue. Results suggested similar koffs for the two 5-OH-DPAT enantiomers at wild-type (WT) D2R, both being slower than the koffs of DA and p-tyramine. Conversely, the kon of (S)-5-OH-DPAT was estimated to be higher than that of (R)-5-OH-DPAT, in agreement with the higher potency of the (S)-enantiomer. Furthermore, S1935.42A mutation lowered the kon of (S)-5-OH-DPAT and reduced the potency difference between the two 5-OH-DPAT enantiomers. Kinetic Kds derived from the koff and kon estimates correlated well with EC50 values for all four compounds across four orders of magnitude, strengthening the notion that our assay captured meaningful information about binding kinetics. The approach presented here may thus prove valuable for characterizing D2R agonist candidate drugs.MDPI202120212021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/47347http://dx.doi.org/10.3390/ijms22084078reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésInt J Mol Sci. 2021;22(8):4078© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10230/473472026-05-29T05:05:01Z
dc.title.none.fl_str_mv Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
title Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
spellingShingle Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
Ågren, Richard
G protein-coupled receptor
Xenopus oocytes
Aminotetralins
Electrophysiology
Molecular dynamics simulation
Phenethylamines
Voltage-clamp
title_short Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
title_full Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
title_fullStr Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
title_full_unstemmed Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
title_sort Dopamine D 2 receptor agonist binding kinetics-role of a conserved serine residue
dc.creator.none.fl_str_mv Ågren, Richard
Stepniewski, Tomasz Maciej, 1988-
Zeberg, Hugo
Selent, Jana
Sahlholm, Kristoffer
author Ågren, Richard
author_facet Ågren, Richard
Stepniewski, Tomasz Maciej, 1988-
Zeberg, Hugo
Selent, Jana
Sahlholm, Kristoffer
author_role author
author2 Stepniewski, Tomasz Maciej, 1988-
Zeberg, Hugo
Selent, Jana
Sahlholm, Kristoffer
author2_role author
author
author
author
dc.subject.none.fl_str_mv G protein-coupled receptor
Xenopus oocytes
Aminotetralins
Electrophysiology
Molecular dynamics simulation
Phenethylamines
Voltage-clamp
topic G protein-coupled receptor
Xenopus oocytes
Aminotetralins
Electrophysiology
Molecular dynamics simulation
Phenethylamines
Voltage-clamp
description The forward (kon) and reverse (koff) rate constants of drug-target interactions have important implications for therapeutic efficacy. Hence, time-resolved assays capable of measuring these binding rate constants may be informative to drug discovery efforts. Here, we used an ion channel activation assay to estimate the kons and koffs of four dopamine D2 receptor (D2R) agonists; dopamine (DA), p-tyramine, (R)- and (S)-5-OH-dipropylaminotetralin (DPAT). We further probed the role of the conserved serine S1935.42 by mutagenesis, taking advantage of the preferential interaction of (S)-, but not (R)-5-OH-DPAT with this residue. Results suggested similar koffs for the two 5-OH-DPAT enantiomers at wild-type (WT) D2R, both being slower than the koffs of DA and p-tyramine. Conversely, the kon of (S)-5-OH-DPAT was estimated to be higher than that of (R)-5-OH-DPAT, in agreement with the higher potency of the (S)-enantiomer. Furthermore, S1935.42A mutation lowered the kon of (S)-5-OH-DPAT and reduced the potency difference between the two 5-OH-DPAT enantiomers. Kinetic Kds derived from the koff and kon estimates correlated well with EC50 values for all four compounds across four orders of magnitude, strengthening the notion that our assay captured meaningful information about binding kinetics. The approach presented here may thus prove valuable for characterizing D2R agonist candidate drugs.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/47347
http://dx.doi.org/10.3390/ijms22084078
url http://hdl.handle.net/10230/47347
http://dx.doi.org/10.3390/ijms22084078
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Int J Mol Sci. 2021;22(8):4078
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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