Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Biol. Chem. 276 (29): 27071-27076

© 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

Plasma Membrane Estrogen Receptors Are Coupled to Endothelial Nitric-oxide Synthase through Galpha i*

Myra H. WyckoffDagger , Ken L. ChamblissDagger , Chieko MineoDagger , Ivan S. YuhannaDagger , Michael E. Mendelsohn§, Susanne M. Mumby, and Philip W. ShaulDagger ||

From the Departments of Dagger  Pediatrics and  Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75390 and the § Molecular Cardiology Research Institute, New England Medical Center and Tufts University School of Medicine, Boston, Massachusetts 02111

Estrogen causes rapid endothelial nitric oxide (NO) production because of the activation of plasma membrane-associated estrogen receptors (ER) coupled to endothelial NO synthase (eNOS). In the present study, we determined the role of G proteins in eNOS activation by estrogen. Estradiol-17beta (E2, 10-8 M) and acetylcholine (10-5 M) caused comparable increases in NOS activity (15 min) in intact endothelial cells that were fully blocked by pertussis toxin (Ptox). In addition, exogenous guanosine 5'-O-(2- thiodiphosphate) inhibited E2-mediated eNOS stimulation in isolated endothelial plasma membranes, and Ptox prevented enzyme activation by E2 in COS-7 cells expressing ERalpha and eNOS. Coimmunoprecipitation studies of plasma membranes from COS-7 cells transfected with ERalpha and specific Galpha proteins demonstrated E2-stimulated interaction between ERalpha and Galpha i but not between ERalpha and either Galpha q or Galpha s; the observed ERalpha -Galpha i interaction was blocked by the ER antagonist ICI 182,780 and by Ptox. E2-stimulated ERalpha -Galpha i interaction was also demonstrable in endothelial cell plasma membranes. Cotransfection of Galpha i into COS-7 cells expressing ERalpha and eNOS yielded a 3-fold increase in E2-mediated eNOS stimulation, whereas cotransfection with a protein regulator of G protein signaling, RGS4, inhibited the E2 response. These findings indicate that eNOS stimulation by E2 requires plasma membrane ERalpha coupling to Galpha i and that activated Galpha i mediates the requisite downstream signaling events. Thus, novel G protein coupling enables a subpopulation of ERalpha to initiate signal transduction at the cell surface. Similar mechanisms may underly the nongenomic actions of other steroid hormones.

* This work was supported by National Institutes of Health Grants HL58888, HL53546, and HD30276 (to P. W. S.), GM50515 (to S. M. M.), and HL56069 and HL59953 (to M. E. M). The project was supported in part by the Lowe Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed. Tel.: 214-648-2015; Fax: 214-648-2481; E-mail:

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

Estrogen Reduces Lipid Content in the Liver Exclusively from Membrane Receptor Signaling.
A. Pedram, M. Razandi, F. O'Mahony, H. Harvey, B. J. Harvey, and E. R. Levin (2013)
Science Signaling 6, ra36
   Abstract »    Full Text »    PDF »
G{alpha}o potentiates estrogen receptor {alpha} activity via the ERK signaling pathway.
M. R. Bratton, J. W. Antoon, B. N. Duong, D. E. Frigo, S. Tilghman, B. M. Collins-Burow, S. Elliott, Y. Tang, L. I. Melnik, L. Lai, et al. (2012)
J. Endocrinol. 214, 45-54
   Abstract »    Full Text »    PDF »
Splice isoform estrogen receptors as integral transmembrane proteins.
K. H. Kim, D. Toomre, and J. R. Bender (2011)
Mol. Biol. Cell 22, 4415-4423
   Abstract »    Full Text »    PDF »
Non-nuclear Estrogen Receptor Signaling in the Endothelium.
Q. Wu, K. Chambliss, M. Umetani, C. Mineo, and P. W. Shaul (2011)
J. Biol. Chem. 286, 14737-14743
   Abstract »    Full Text »    PDF »
Estrogen Actions in the Brain and the Basis for Differential Action in Men and Women: A Case for Sex-Specific Medicines.
G. E. Gillies and S. McArthur (2010)
Pharmacol. Rev. 62, 155-198
   Abstract »    Full Text »    PDF »
Identification of a Membrane Estrogen Receptor in Zebrafish with Homology to Mammalian GPER and Its High Expression in Early Germ Cells of the Testis.
X. Liu, P. Zhu, K. W.Y. Sham, J. M.L. Yuen, C. Xie, Y. Zhang, Y. Liu, S. Li, X. Huang, C. H.K. Cheng, et al. (2009)
Biol Reprod 80, 1253-1261
   Abstract »    Full Text »    PDF »
17{beta}-Estradiol-Mediated Neuroprotection and ERK Activation Require a Pertussis Toxin-Sensitive Mechanism Involving GRK2 and {beta}-Arrestin-1.
R. Dominguez, E. Hu, M. Zhou, and M. Baudry (2009)
J. Neurosci. 29, 4228-4238
   Abstract »    Full Text »    PDF »
Glucocorticoids Regulate Glutamate and GABA Synapse-Specific Retrograde Transmission via Divergent Nongenomic Signaling Pathways.
S. Di, M. M. Maxson, A. Franco, and J. G. Tasker (2009)
J. Neurosci. 29, 393-401
   Abstract »    Full Text »    PDF »
Resveratrol stimulates nitric oxide production by increasing estrogen receptor {alpha}-Src-caveolin-1 interaction and phosphorylation in human umbilical vein endothelial cells.
C. M. Klinge, N. S. Wickramasinghe, M. M. Ivanova, and S. M. Dougherty (2008)
FASEB J 22, 2185-2197
   Abstract »    Full Text »    PDF »
EF Domains Are Sufficient for Nongenomic Mineralocorticoid Receptor Actions.
C. Grossmann, R. Freudinger, S. Mildenberger, B. Husse, and M. Gekle (2008)
J. Biol. Chem. 283, 7109-7116
   Abstract »    Full Text »    PDF »
Anti-hepatitis C Virus Activity of Tamoxifen Reveals the Functional Association of Estrogen Receptor with Viral RNA Polymerase NS5B.
K. Watashi, D. Inoue, M. Hijikata, K. Goto, H. H. Aly, and K. Shimotohno (2007)
J. Biol. Chem. 282, 32765-32772
   Abstract »    Full Text »    PDF »
Nongenomic {beta} Estrogen Receptors Enhance {beta}1 Adrenergic Signaling Induced by the Nicotine-Derived Carcinogen 4-(Methylnitrosamino)-1-(3-Pyridyl)-1-Butanone in Human Small Airway Epithelial Cells.
M. Majidi, H. A. Al-Wadei, T. Takahashi, and H. M. Schuller (2007)
Cancer Res. 67, 6863-6871
   Abstract »    Full Text »    PDF »
Defective Insulin and Acetylcholine Induction of Endothelial Cell-Nitric Oxide Synthase Through Insulin Receptor Substrate/Akt Signaling Pathway in Aorta of Obese Rats.
H. G. Zecchin, F. B.M. Priviero, C. T. Souza, K. G. Zecchin, P. O. Prada, J. B.C. Carvalheira, L. A. Velloso, E. Antunes, and M. J.A. Saad (2007)
Diabetes 56, 1014-1024
   Abstract »    Full Text »    PDF »
Estrogen Mobilizes a Subset of Estrogen Receptor-{alpha}-Immunoreactive Vesicles in Inhibitory Presynaptic Boutons in Hippocampal CA1.
S. A. Hart, M. A. Snyder, T. Smejkalova, and C. S. Woolley (2007)
J. Neurosci. 27, 2102-2111
   Abstract »    Full Text »    PDF »
Role of receptor complexes in the extranuclear actions of estrogen receptor {alpha} in breast cancer.
R. X-D Song, P. Fan, W. Yue, Y. Chen, and R. J Santen (2006)
Endocr. Relat. Cancer 13, S3-S13
   Abstract »    Full Text »    PDF »
Oxygen alters caveolin-1 and nitric oxide synthase-3 functions in ovine fetal and neonatal lung microvascular endothelial cells.
T. A. John, B. O. Ibe, and J. Usha Raj (2006)
Am J Physiol Lung Cell Mol Physiol 291, L1079-L1093
   Abstract »    Full Text »    PDF »
S. Joy, R. C. M. Siow, D. J. Rowlands, M. Becker, A. W. Wyatt, P. I. Aaronson, C. W. Coen, I. Kallo, R. Jacob, and G. E. Mann (2006)
J. Biol. Chem. 281, 27335-27345
   Abstract »    Full Text »    PDF »
Membrane Initiated Estrogen Signaling in Breast Cancer.
R. X.-D. Song and R. J. Santen (2006)
Biol Reprod 75, 9-16
   Abstract »    Full Text »    PDF »
Estradiol-17beta stimulates proliferation of mouse embryonic stem cells: involvement of MAPKs and CDKs as well as protooncogenes.
H. J. Han, J. S. Heo, and Y. J. Lee (2006)
Am J Physiol Cell Physiol 290, C1067-C1075
   Abstract »    Full Text »    PDF »
Circulating cardiovascular disease risk factors and signaling in endothelial cell caveolae.
C. Mineo and P. W. Shaul (2006)
Cardiovasc Res 70, 31-41
   Abstract »    Full Text »    PDF »
Role of estrogen receptor (ER) {alpha} in insulin-like growth factor (IGF)-I-induced responses in MCF-7 breast cancer cells.
S Zhang, X Li, R Burghardt, R Smith III, and S H Safe (2005)
J. Mol. Endocrinol. 35, 433-447
   Abstract »    Full Text »    PDF »
Microvascular effects of corticotropin-releasing hormone in human skin vary in relation to estrogen concentration during the menstrual cycle.
V L Clifton, R Crompton, M A Read, P G Gibson, R Smith, and I M R Wright (2005)
J. Endocrinol. 186, 69-76
   Abstract »    Full Text »    PDF »
Rapid, Estrogen Receptor-Mediated Signaling: Why Is the Endothelium So Special?.
K. H. Kim and J. R. Bender (2005)
Sci. STKE 2005, pe28
   Abstract »    Full Text »    PDF »
Estradiol Activates Group I and II Metabotropic Glutamate Receptor Signaling, Leading to Opposing Influences on cAMP Response Element-Binding Protein.
M. I. Boulware, J. P. Weick, B. R. Becklund, S. P. Kuo, R. D. Groth, and P. G. Mermelstein (2005)
J. Neurosci. 25, 5066-5078
   Abstract »    Full Text »    PDF »
Nongenotropic, Anti-Apoptotic Signaling of 1{alpha},25(OH)2-Vitamin D3 and Analogs through the Ligand Binding Domain of the Vitamin D Receptor in Osteoblasts and Osteocytes: MEDIATION BY Src, PHOSPHATIDYLINOSITOL 3-, AND JNK KINASES.
A. M. Vertino, C. M. Bula, J.-R. Chen, M. Almeida, L. Han, T. Bellido, S. Kousteni, A. W. Norman, and S. C. Manolagas (2005)
J. Biol. Chem. 280, 14130-14137
   Abstract »    Full Text »    PDF »
Resveratrol and Estradiol Rapidly Activate MAPK Signaling through Estrogen Receptors {alpha} and {beta} in Endothelial Cells.
C. M. Klinge, K. A. Blankenship, K. E. Risinger, S. Bhatnagar, E. L. Noisin, W. K. Sumanasekera, L. Zhao, D. M. Brey, and R. S. Keynton (2005)
J. Biol. Chem. 280, 7460-7468
   Abstract »    Full Text »    PDF »
Rapid nitric oxide-mediated S-nitrosylation of estrogen receptor: Regulation of estrogen-dependent gene transcription.
H. J. Garban, D. C. Marquez-Garban, R. J. Pietras, and L. J. Ignarro (2005)
PNAS 102, 2632-2636
   Abstract »    Full Text »    PDF »
Striatin assembles a membrane signaling complex necessary for rapid, nongenomic activation of endothelial NO synthase by estrogen receptor {alpha}.
Q. Lu, D. C. Pallas, H. K. Surks, W. E. Baur, M. E. Mendelsohn, and R. H. Karas (2004)
PNAS 101, 17126-17131
   Abstract »    Full Text »    PDF »
The G Protein-coupled Receptor GPR30 Mediates c-fos Up-regulation by 17{beta}-Estradiol and Phytoestrogens in Breast Cancer Cells.
M. Maggiolini, A. Vivacqua, G. Fasanella, A. G. Recchia, D. Sisci, V. Pezzi, D. Montanaro, A. M. Musti, D. Picard, and S. Ando (2004)
J. Biol. Chem. 279, 27008-27016
   Abstract »    Full Text »    PDF »
Ribozyme knockdown functionally links a 1,25(OH)2D3 membrane binding protein (1,25D3-MARRS) and phosphate uptake in intestinal cells.
I. Nemere, M. C. Farach-Carson, B. Rohe, T. M. Sterling, A. W. Norman, B. D. Boyan, and S. E. Safford (2004)
PNAS 101, 7392-7397
   Abstract »    Full Text »    PDF »
Thombospondin-1 Disrupts Estrogen-Induced Endothelial Cell Proliferation and Migration and Its Expression Is Suppressed by Estradiol.
K. Sengupta, S. Banerjee, N. K. Saxena, and S. K. Banerjee (2004)
Mol. Cancer Res. 2, 150-158
   Abstract »    Full Text »    PDF »
Functional Melatonin Receptors in Rat Ovaries at Various Stages of the Estrous Cycle.
J. M. Soares Jr., M. I. Masana, C. Ersahin, and M. L. Dubocovich (2003)
J. Pharmacol. Exp. Ther. 306, 694-702
   Abstract »    Full Text »    PDF »
Nongenomic Steroid Action: Controversies, Questions, and Answers.
Physiol Rev 83, 965-1016
   Abstract »    Full Text »    PDF »
Impaired vasomodulation is associated with reduced neuronal nitric oxide synthase in skeletal muscle of ovariectomized rats.
P. J Fadel, W. Zhao, and G. D Thomas (2003)
J. Physiol. 549, 243-253
   Abstract »    Full Text »    PDF »
Estrogen Receptor, Cyclic Adenosine Monophosphate, and Protein Kinase A Are Involved in the Nongenomic Pathway by Which Estradiol Accelerates Oviductal Oocyte Transport in Cyclic Rats.
P. A. Orihuela, A. Parada-Bustamante, P. P. Cortes, C. Gatica, and H. B. Croxatto (2003)
Biol Reprod 68, 1225-1231
   Abstract »    Full Text »    PDF »
Identification of a Structural Determinant Necessary for the Localization and Function of Estrogen Receptor {alpha} at the Plasma Membrane.
M. Razandi, G. Alton, A. Pedram, S. Ghonshani, P. Webb, and E. R. Levin (2003)
Mol. Cell. Biol. 23, 1633-1646
   Abstract »    Full Text »    PDF »
Possible mechanisms underlying pregnancy-induced changes in uterine artery endothelial function.
I. M. Bird, L. Zhang, and R. R. Magness (2003)
Am J Physiol Regulatory Integrative Comp Physiol 284, R245-R258
   Abstract »    Full Text »    PDF »
Proximal Events in Signaling by Plasma Membrane Estrogen Receptors.
M. Razandi, A. Pedram, S. T. Park, and E. R. Levin (2003)
J. Biol. Chem. 278, 2701-2712
   Abstract »    Full Text »    PDF »
Integration of the Non-genomic and Genomic Actions of Estrogen. MEMBRANE-INITIATED SIGNALING BY STEROID TO TRANSCRIPTION AND CELL BIOLOGY.
A. Pedram, M. Razandi, M. Aitkenhead, C. C. W. Hughes, and E. R. Levin (2002)
J. Biol. Chem. 277, 50768-50775
   Abstract »    Full Text »    PDF »
Nonnuclear Actions of Estrogen.
K. J. Ho and J. K. Liao (2002)
Arterioscler Thromb Vasc Biol 22, 1952-1961
   Abstract »    Full Text »    PDF »
ER-X: A Novel, Plasma Membrane-Associated, Putative Estrogen Receptor That Is Regulated during Development and after Ischemic Brain Injury.
C. D. Toran-Allerand, X. Guan, N. J. MacLusky, T. L. Horvath, S. Diano, M. Singh, E. S. Connolly Jr, I. S. Nethrapalli, and A. A. Tinnikov (2002)
J. Neurosci. 22, 8391-8401
   Abstract »    Full Text »    PDF »
Dehydroepiandrosterone Activates Endothelial Cell Nitric-oxide Synthase by a Specific Plasma Membrane Receptor Coupled to Galpha i2,3.
D. Liu and J. S. Dillon (2002)
J. Biol. Chem. 277, 21379-21388
   Abstract »    Full Text »    PDF »
Nongenomic Regulation of the Kinetics of Exocytosis by Estrogens.
J. D. Machado, C. Alonso, A. Morales, J. F. Gomez, and R. Borges (2002)
J. Pharmacol. Exp. Ther. 301, 631-637
   Abstract »    Full Text »    PDF »
Early membrane estrogenic effects required for full expression of slower genomic actions in a nerve cell line.
N. Vasudevan, L.-M. Kow, and D. W. Pfaff (2001)
PNAS 98, 12267-12271
   Abstract »    Full Text »    PDF »
Early membrane estrogenic effects required for full expression of slower genomic actions in a nerve cell line.
N. Vasudevan, L.-M. Kow, and D. W. Pfaff (2001)
PNAS 98, 12267-12271
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882