Differential ligand binding affinities of human estrogen receptor-α isoforms.

Rapid 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-α4...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Amanda H Y Lin, Rachel W S Li, Eva Y W Ho, George P H Leung, Susan W S Leung, Paul M Vanhoutte, Ricky Y K Man
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/92b20d56ea004f4a8e31b58a69038324
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:92b20d56ea004f4a8e31b58a69038324
record_format	dspace
spelling	oai:doaj.org-article:92b20d56ea004f4a8e31b58a690383242021-11-18T07:47:18ZDifferential ligand binding affinities of human estrogen receptor-α isoforms.1932-620310.1371/journal.pone.0063199https://doaj.org/article/92b20d56ea004f4a8e31b58a690383242013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23646196/?tool=EBIhttps://doaj.org/toc/1932-6203Rapid 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.Amanda H Y LinRachel W S LiEva Y W HoGeorge P H LeungSusan W S LeungPaul M VanhoutteRicky Y K ManPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 4, p e63199 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Amanda H Y Lin Rachel W S Li Eva Y W Ho George P H Leung Susan W S Leung Paul M Vanhoutte Ricky Y K Man Differential ligand binding affinities of human estrogen receptor-α isoforms.
description	Rapid 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.
format	article
author	Amanda H Y Lin Rachel W S Li Eva Y W Ho George P H Leung Susan W S Leung Paul M Vanhoutte Ricky Y K Man
author_facet	Amanda H Y Lin Rachel W S Li Eva Y W Ho George P H Leung Susan W S Leung Paul M Vanhoutte Ricky Y K Man
author_sort	Amanda H Y Lin
title	Differential ligand binding affinities of human estrogen receptor-α isoforms.
title_short	Differential ligand binding affinities of human estrogen receptor-α isoforms.
title_full	Differential ligand binding affinities of human estrogen receptor-α isoforms.
title_fullStr	Differential ligand binding affinities of human estrogen receptor-α isoforms.
title_full_unstemmed	Differential ligand binding affinities of human estrogen receptor-α isoforms.
title_sort	differential ligand binding affinities of human estrogen receptor-α isoforms.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/92b20d56ea004f4a8e31b58a69038324
work_keys_str_mv	AT amandahylin differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT rachelwsli differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT evaywho differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT georgephleung differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT susanwsleung differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT paulmvanhoutte differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms AT rickyykman differentialligandbindingaffinitiesofhumanestrogenreceptoraisoforms
_version_	1718422987580899328

Differential ligand binding affinities of human estrogen receptor-α isoforms.

Ejemplares similares