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dc.contributor.authorThompson, Andrewen
dc.contributor.authorMetzger, Sen
dc.contributor.authorLochner, Men
dc.contributor.authorRuepp, M-Den
dc.date.accessioned2017-03-28T16:00:42Z
dc.date.available2017-03-28T16:00:42Z
dc.date.issued2017-04-01en
dc.identifier.issn0028-3908
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/263300
dc.description.abstractEpibatidine is an alkaloid toxin that binds with high affinity to nicotinic and muscarinic acetylcholine receptors, and has been extensively used as a research tool. To examine binding interactions at the nicotinic receptor, it has been co-crystallised with the structural homologue acetylcholine binding protein (AChBP; PDB ID 2BYQ), and with an AChBP chimaera (3SQ6) that shares 64% sequence identity with the α7 nACh receptor. However, the binding orientations revealed by AChBP co-crystal structures may not precisely represent their receptor homologues and experimental evidence is needed to verify the ligand poses. Here we identify potential binding site interactions between epibatidine and AChBP residues, and substitute equivalent positions in the α7 nACh receptor. The effects of these are probed by [$^{3}$H]epibatidine binding following the expression α7 nACh receptor cysteine mutants in HEK 293 cells. Of the sixteen mutants created, the affinity of epibatidine was unaffected by the substitutions Q55C, L106C, L116C, T146C, D160C and S162C, reduced by C186A and C187A, increased by Q114C and S144C, and abolished by W53C, Y91C, N104C, W145C, Y184C and Y191C. These results are consistent with the predicted orientations in AChBP and suggest that epibatidine is likely to occupy a similar location at α7 nACh receptors. We speculate that steric constraints placed upon the C-5 position of the pyridine ring in 3SQ6 may account for the relatively poor affinities of epibatidine derivatives that are substituted at this position.
dc.description.sponsorshipThis work was funded by the British Heart Foundation [PG/13/39/3029 to AJT], HOLCIM Stiftung zur Förderung der wissenschaftlichen Fortbildung [to M-DR], the NOMIS Foundation [to M-DR], and the Swiss National Science Foundation [SNSF professorship PP00P2_146321/1 to ML].
dc.languageengen
dc.language.isoenen
dc.publisherElsevier
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rightsAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectcys-loopen
dc.subjection channelen
dc.subjectradioliganden
dc.subjectepibatidineen
dc.subjectagonisten
dc.subjectα7 nAChen
dc.subjectnicotinicen
dc.subjectreceptoren
dc.subjectAChBPen
dc.titleThe binding orientation of epibatidine at α7 nACh receptorsen
dc.typeArticle
prism.endingPage428
prism.publicationDate2017en
prism.publicationNameNeuropharmacologyen
prism.startingPage421
prism.volume116en
dc.identifier.doi10.17863/CAM.8635
dcterms.dateAccepted2017-01-11en
rioxxterms.versionofrecord10.1016/j.neuropharm.2017.01.008en
rioxxterms.versionVoRen
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/en
rioxxterms.licenseref.startdate2017-04-01en
dc.contributor.orcidThompson, Andrew [0000-0002-7046-6792]
dc.identifier.eissn1873-7064
rioxxterms.typeJournal Article/Reviewen
pubs.funder-project-idBritish Heart Foundation (PG/13/39/30293)
cam.issuedOnline2017-01-12en
cam.orpheus.successThu Jan 30 12:56:48 GMT 2020 - The item has an open VoR version.*
rioxxterms.freetoread.startdate2100-01-01


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International