Ligand with two modes of interaction with the dopamine D2 receptor-an induced-fit mechanism of insurmountable antagonism

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• dc.contributor.author Ågren, Richard
• dc.contributor.author Zeberg, Hugo
• dc.contributor.author Stepniewski, Tomasz Maciej, 1988-
• dc.contributor.author Free, R. Benjamin
• dc.contributor.author Reilly, Sean W.
• dc.contributor.author Luedtke, Robert R.
• dc.contributor.author Århem, Peter
• dc.contributor.author Ciruela, Francisco
• dc.contributor.author Sibley, David R.
• dc.contributor.author Mach, Robert H.
• dc.contributor.author Selent, Jana
• dc.contributor.author Nilsson, Johanna
• dc.contributor.author Sahlholm, Kristoffer
• dc.date.accessioned 2020-11-03T06:37:20Z
• dc.date.available 2020-11-03T06:37:20Z
• dc.date.issued 2020
• dc.description.abstract A solid understanding of the mechanisms governing ligand binding is crucial for rational design of therapeutics targeting the dopamine D2 receptor (D2R). Here, we use G protein-coupled inward rectifier potassium (GIRK) channel activation in Xenopus oocytes to measure the kinetics of D2R antagonism by a series of aripiprazole analogues, as well as the recovery of dopamine (DA) responsivity upon washout. The aripiprazole analogues comprise an orthosteric and a secondary pharmacophore and differ by the length of the saturated carbon linker joining these two pharmacophores. Two compounds containing 3- and 5-carbon linkers allowed for a similar extent of recovery from antagonism in the presence of 1 or 100 μM DA (>25 and >90% of control, respectively), whereas recovery was less prominent (∼20%) upon washout of the 4-carbon linker compound, SV-III-130, both with 1 and 100 μM DA. Prolonging the coincubation time with SV-III-130 further diminished recovery. Curve-shift experiments were consistent with competition between SV-III-130 and DA. Two mutations in the secondary binding pocket (V91A and E95A) of D2R decreased antagonistic potency and increased recovery from SV-III-130 antagonism, whereas a third mutation (L94A) only increased recovery. Our results suggest that the secondary binding pocket influences recovery from inhibition by the studied aripiprazole analogues. We propose a mechanism, supported by in silico modeling, whereby SV-III-130 initially binds reversibly to the D2R, after which the drug-receptor complex undergoes a slow transition to a second ligand-bound state, which is dependent on secondary binding pocket integrity and irreversible during the time frame of our experiments.
• dc.description.sponsorship This study was supported by grants from Stiftelsen Lars Hiertas Minne, Åhlénstiftelsen, Karolinska Institutet Funds and Magnus Bergvalls stiftelse (to K.S.). K.S. received postdoctoral grants from the Swedish Brain Foundation and the Swedish Society of Medicine and is currently a fellow at the Wallenberg Center for Molecular Medicine at Umeå University. T.M.S. acknowledges financial support from the National Science Centre of Poland, project number 2017/27/N/NZ2/02571. J.S. received financial support from the Instituto de Salud Carlos III FEDER (PI15/00460 & PI18/00094) and the NEURON-ERANET (AC18/00030). F.C. was supported by FEDER/Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación (SAF2017-87349-R) and the Catalan government (2017 SGR 1604). R.B.F. and D.R.S. are supported by the National Institutes of Health, National Institute of Neurological Disorders and Stroke-Intramural Research Program (ZIA NS002263). We would like to thank Brittney Miller and Martin Dalefield for excellent technical assistance.
• dc.format.mimetype application/pdf
• dc.identifier.citation Ågren R, Zeberg H, Stępniewski TM, Free RB, Reilly SW, Luedtke RR, Århem P, Ciruela F, Sibley DR, Mach RH, Selent J, Nilsson J, Sahlholm K. Ligand with two modes of interaction with the dopamine D2 receptor-an induced-fit mechanism of insurmountable antagonism. ACS Chem Neurosci. 2020; 11(19):3130-43. DOI: 10.1021/acschemneuro.0c00477
• dc.identifier.doi http://dx.doi.org/10.1021/acschemneuro.0c00477
• dc.identifier.issn 1948-7193
• dc.identifier.uri http://hdl.handle.net/10230/45642
• dc.language.iso eng
• dc.publisher American Chemical Society (ACS)
• dc.relation.ispartof ACS Chem Neurosci. 2020; 11(19):3130-43
• dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/SAF2017-87349-R
• dc.rights © 2020 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium,provided the author and source are cited.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri https://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword HEK cells
• dc.subject.keyword PET scan
• dc.subject.keyword Xenopus oocytes
• dc.subject.keyword Antipsychotics
• dc.subject.keyword Arrestin
• dc.subject.keyword Competitive binding
• dc.subject.keyword Drug kinetics
• dc.subject.keyword Molecular dynamics simulation
• dc.title Ligand with two modes of interaction with the dopamine D2 receptor-an induced-fit mechanism of insurmountable antagonism
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion