Advanced

G Protein-Coupled Estrogen Receptor 1 (GPER1)/GPR30 Localizes in the Plasma Membrane and Trafficks Intracellularly on Cytokeratin Intermediate Filaments.

Sandén, Caroline LU ; Broselid, Stefan LU ; Cornmark, Louise LU ; Andersson, Krister LU ; Daszkiewicz-Nilsson, Joanna LU ; Mårtensson, Ulrika LU ; Olde, Björn LU and Leeb-Lundberg, Fredrik LU (2011) In Molecular Pharmacology 79(3). p.400-410
Abstract
GPR30, or G protein-coupled estrogen receptor 1 (GPER1), was recently introduced as a membrane estrogen receptor and a candidate cancer biomarker and therapeutic target. However, several questions surround the subcellular localization and signaling of this receptor. In native cells, including mouse myoblast C(2)C(12) cells, Madine-Darby canine kidney (MDCK) epithelial cells, and human ductal breast epithelial tumor T47-D cells, G-1, a GPER1 agonist, and 17β-estradiol (E2) stimulated GPER1-dependent cAMP production, a defined plasma membrane (PM) event, and recruitment of β-arrestin2 to the PM. Staining of fixed and live cells showed that GPER1 was localized both in the PM and on intracellular structures. One such intracellular structure... (More)
GPR30, or G protein-coupled estrogen receptor 1 (GPER1), was recently introduced as a membrane estrogen receptor and a candidate cancer biomarker and therapeutic target. However, several questions surround the subcellular localization and signaling of this receptor. In native cells, including mouse myoblast C(2)C(12) cells, Madine-Darby canine kidney (MDCK) epithelial cells, and human ductal breast epithelial tumor T47-D cells, G-1, a GPER1 agonist, and 17β-estradiol (E2) stimulated GPER1-dependent cAMP production, a defined plasma membrane (PM) event, and recruitment of β-arrestin2 to the PM. Staining of fixed and live cells showed that GPER1 was localized both in the PM and on intracellular structures. One such intracellular structure was identified as cytokeratin (CK) intermediate filaments, including those composed of CK7 and CK8, but apparently not endoplasmatic reticulum (ER), Golgi, or microtubules. Reciprocal co-immunoprecipitation of GPER1 and CKs confirmed an association of these proteins. Live staining also showed that the PM receptors constitutively internalize apparently to reach CK filaments. Receptor localization was supported using FLAG- and HA-tagged GPER1. We conclude that GPER1-mediated stimulation of cAMP production and β-arrestin2 recruitment occur in the PM. Furthermore, the PM receptors constitutively internalize and localize intracellularly on CK. This is the first observation that a G protein-coupled receptor (GPCR) is capable of associating with intermediate filaments, which may be important for GPER1 regulation in epithelial cells and the relationship of this receptor to cancer. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Molecular Pharmacology
volume
79
issue
3
pages
400 - 410
publisher
American Society for Pharmacology and Experimental Therapeutics
external identifiers
  • wos:000287481800008
  • pmid:21149639
  • scopus:79951987933
ISSN
1521-0111
DOI
10.1124/mol.110.069500
language
English
LU publication?
yes
id
982b5ce6-80c8-4149-8138-28d265d09f44 (old id 1756512)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/21149639?dopt=Abstract
date added to LUP
2011-01-03 15:20:48
date last changed
2017-11-12 03:04:24
@article{982b5ce6-80c8-4149-8138-28d265d09f44,
  abstract     = {GPR30, or G protein-coupled estrogen receptor 1 (GPER1), was recently introduced as a membrane estrogen receptor and a candidate cancer biomarker and therapeutic target. However, several questions surround the subcellular localization and signaling of this receptor. In native cells, including mouse myoblast C(2)C(12) cells, Madine-Darby canine kidney (MDCK) epithelial cells, and human ductal breast epithelial tumor T47-D cells, G-1, a GPER1 agonist, and 17β-estradiol (E2) stimulated GPER1-dependent cAMP production, a defined plasma membrane (PM) event, and recruitment of β-arrestin2 to the PM. Staining of fixed and live cells showed that GPER1 was localized both in the PM and on intracellular structures. One such intracellular structure was identified as cytokeratin (CK) intermediate filaments, including those composed of CK7 and CK8, but apparently not endoplasmatic reticulum (ER), Golgi, or microtubules. Reciprocal co-immunoprecipitation of GPER1 and CKs confirmed an association of these proteins. Live staining also showed that the PM receptors constitutively internalize apparently to reach CK filaments. Receptor localization was supported using FLAG- and HA-tagged GPER1. We conclude that GPER1-mediated stimulation of cAMP production and β-arrestin2 recruitment occur in the PM. Furthermore, the PM receptors constitutively internalize and localize intracellularly on CK. This is the first observation that a G protein-coupled receptor (GPCR) is capable of associating with intermediate filaments, which may be important for GPER1 regulation in epithelial cells and the relationship of this receptor to cancer.},
  author       = {Sandén, Caroline and Broselid, Stefan and Cornmark, Louise and Andersson, Krister and Daszkiewicz-Nilsson, Joanna and Mårtensson, Ulrika and Olde, Björn and Leeb-Lundberg, Fredrik},
  issn         = {1521-0111},
  language     = {eng},
  number       = {3},
  pages        = {400--410},
  publisher    = {American Society for Pharmacology and Experimental Therapeutics},
  series       = {Molecular Pharmacology},
  title        = {G Protein-Coupled Estrogen Receptor 1 (GPER1)/GPR30 Localizes in the Plasma Membrane and Trafficks Intracellularly on Cytokeratin Intermediate Filaments.},
  url          = {http://dx.doi.org/10.1124/mol.110.069500},
  volume       = {79},
  year         = {2011},
}