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Article: Differential ligand binding affinities of human estrogen receptor-α isoforms

TitleDifferential ligand binding affinities of human estrogen receptor-α isoforms
Authors
Issue Date2013
Citation
PLOS One, 2013, v. 8, p. e63199 How to Cite?
AbstractRapid non-genomic effects of 17β-estradiol are elicited by the activation of different estrogen receptor-α isoforms. Presence of surface binding sites for estrogen have been identified in cells transfected with full-length estrogen receptor-α66 (ER66) and the truncated isoforms, estrogen receptor-α46 (ER46) and estrogen receptor-α36 (ER36). However, the binding affinities of the membrane estrogen receptors (mERs) remain unknown due to the difficulty of developing of stable mER-transfected cell lines with sufficient mER density, which has largely hampered biochemical binding studies. The present study utilized cell-free expression systems to determine the binding affinities of 17β-estradiol to mERs, and the relationship among palmitoylation, membrane insertion and binding affinities. Saturation binding assays of human mERs revealed that [3H]-17β-estradiol bound ER66 and ER46 with Kd values of 68.81 and 60.72 pM, respectively, whereas ER36 displayed no specific binding within the tested concentration range. Inhibition of palmitoylation or removal of the nanolipoprotein particles, used as membrane substitute, reduced the binding affinities of ER66 and ER46 to 17β-estradiol. Moreover, ER66 and ER46 bound differentially with some estrogen receptor agonists and antagonists, and phytoestrogens. In particular, the classical estrogen receptor antagonist, ICI 182,780, had a higher affinity for ER66 than ER46. In summary, the present study defines the binding affinities for human estrogen receptor-α isoforms, and demonstrates that ER66 and ER46 show characteristics of mERs. The present data also indicates that palmitoylation and membrane insertion of mERs are important for proper receptor conformation allowing 17β-estradiol binding. The differential binding of ER66 and ER46 with certain compounds substantiates the prospect of developing mER-selective drugs.
Persistent Identifierhttp://hdl.handle.net/10722/186131
PubMed Central ID

 

DC FieldValueLanguage
dc.contributor.authorLIN, HYAen_US
dc.contributor.authorLi, WSRen_US
dc.contributor.authorHo, YWen_US
dc.contributor.authorLeung, GPHen_US
dc.contributor.authorLeung, SWSen_US
dc.contributor.authorVanhoutte, PMGRen_US
dc.contributor.authorMan, RYKen_US
dc.date.accessioned2013-08-20T11:56:22Z-
dc.date.available2013-08-20T11:56:22Z-
dc.date.issued2013en_US
dc.identifier.citationPLOS One, 2013, v. 8, p. e63199en_US
dc.identifier.urihttp://hdl.handle.net/10722/186131-
dc.description.abstractRapid non-genomic effects of 17β-estradiol are elicited by the activation of different estrogen receptor-α isoforms. Presence of surface binding sites for estrogen have been identified in cells transfected with full-length estrogen receptor-α66 (ER66) and the truncated isoforms, estrogen receptor-α46 (ER46) and estrogen receptor-α36 (ER36). However, the binding affinities of the membrane estrogen receptors (mERs) remain unknown due to the difficulty of developing of stable mER-transfected cell lines with sufficient mER density, which has largely hampered biochemical binding studies. The present study utilized cell-free expression systems to determine the binding affinities of 17β-estradiol to mERs, and the relationship among palmitoylation, membrane insertion and binding affinities. Saturation binding assays of human mERs revealed that [3H]-17β-estradiol bound ER66 and ER46 with Kd values of 68.81 and 60.72 pM, respectively, whereas ER36 displayed no specific binding within the tested concentration range. Inhibition of palmitoylation or removal of the nanolipoprotein particles, used as membrane substitute, reduced the binding affinities of ER66 and ER46 to 17β-estradiol. Moreover, ER66 and ER46 bound differentially with some estrogen receptor agonists and antagonists, and phytoestrogens. In particular, the classical estrogen receptor antagonist, ICI 182,780, had a higher affinity for ER66 than ER46. In summary, the present study defines the binding affinities for human estrogen receptor-α isoforms, and demonstrates that ER66 and ER46 show characteristics of mERs. The present data also indicates that palmitoylation and membrane insertion of mERs are important for proper receptor conformation allowing 17β-estradiol binding. The differential binding of ER66 and ER46 with certain compounds substantiates the prospect of developing mER-selective drugs.-
dc.languageengen_US
dc.relation.ispartofPLOS Oneen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleDifferential ligand binding affinities of human estrogen receptor-α isoformsen_US
dc.typeArticleen_US
dc.identifier.emailLi, WSR: h0594069@hkusua.hku.hken_US
dc.identifier.emailHo, YW: eywho@graduate.hku.hken_US
dc.identifier.emailLeung, GPH: gphleung@hkucc.hku.hken_US
dc.identifier.emailLeung, SWS: swsleung@hku.hken_US
dc.identifier.emailVanhoutte, PMGR: vanhoutt@hku.hken_US
dc.identifier.emailMan, RYK: rykman@hkucc.hku.hken_US
dc.identifier.authorityLeung, GPH=rp00234en_US
dc.identifier.authorityLeung, SWS=rp00235en_US
dc.identifier.authorityVanhoutte, PMGR=rp00238en_US
dc.identifier.authorityMan, RYK=rp00236en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1371/journal.pone.0063199-
dc.identifier.pmcidPMC3639985-
dc.identifier.hkuros217587en_US
dc.identifier.volume8en_US
dc.identifier.spagee63199en_US
dc.identifier.epagee63199en_US

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