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The FASEB Journal, Vol 9, 404-410, Copyright © 1995 by The Federation of American Societies for Experimental Biology

RESEARCH COMMUNICATIONS

Membrane estrogen receptors identified by multiple antibody labeling and impeded-ligand binding

TC Pappas, B Gametchu and CS Watson
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77555-0645.

GH3/B6 rat pituitary tumor cells exhibit rapid prolactin release (within 5 min) when treated with nanomolar amounts of estrogen. However, the putative protein mediator of this nongenomic action has not been described. Using antibodies directed against a peptide representing the hinge region of the intracellular estrogen receptor (iER), we have demonstrated that these cells contain a membrane ER (mER). We now report that confocal scanning laser microscopy of cells labeled live with the anti-peptide antibody further supports a membrane localization of ER. The monoclonal antibodies H226 and H222 and a polyclonal antibody, ER21, each recognizing a unique epitope on iER (NH2 terminal to the DNA-binding region, within the steroid binding region, and the NH2-terminal end, respectively), also immunohistochemically label membrane proteins of immuno-selected GH3/B6 cells. These cells also specifically bind a fluorescent estrogen-BSA conjugate. Coincubation of cells with anti-ER antibody and the fluorescent estrogen-BSA conjugate reveals that these labels colocalize on cells. These results suggest that mER may be structurally similar to iER.


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J. Neurosci., February 15, 1999; 19(4): 1179 - 1188.
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M. Razandi, A. Pedram, G. L. Greene, and E. R. Levin
Cell Membrane and Nuclear Estrogen Receptors (ERs) Originate from a Single Transcript: Studies of ER{alpha} and ER{beta} Expressed in Chinese Hamster Ovary Cells
Mol. Endocrinol., February 1, 1999; 13(2): 307 - 319.
[Abstract] [Full Text]

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Q. Gu, K. S. Korach, and R. L. Moss
Rapid Action of 17{beta}-Estradiol on Kainate-Induced Currents in Hippocampal Neurons Lacking Intracellular Estrogen Receptors
Endocrinology, February 1, 1999; 140(2): 660 - 666.
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W. P. M. benten, M. Lieberherr, G. Giese, C. Wrehlke, O. Stamm, C. E. Sekeris, H. Mossmann, and F. Wunderlich
Functional testosterone receptors in plasma membranes of T cells
FASEB J, January 1, 1999; 13(1): 123 - 133.
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S. Jovanovic, A. Jovanovic, W. K. Shen, and A. Terzic
Protective action of 17{beta}-estradiol in cardiac cells: implications for hyperkalemic cardioplegia
Ann. Thorac. Surg., November 1, 1998; 66(5): 1658 - 1661.
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A. Nadal, J. M. Rovira, O. Laribi, T. Leon-quinto, E. Andreu, C. Ripoll, and B. Soria
Rapid insulinotropic effect of 17฿-estradiol via a plasma membrane receptor
FASEB J, October 1, 1998; 12(13): 1341 - 1348.
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M. Christ and M. Wehling
Cardiovascular steroid actions: swift swallows or sluggish snails?
Cardiovasc Res, October 1, 1998; 40(1): 34 - 44.
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D. O. Ruehlmann, J. R. Steinert, M. A. Valverde, R. Jacob, and G. E. Mann
Environmental estrogenic pollutants induce acute vascular relaxation by inhibiting L-type Ca2+ channels in smooth muscle cells
FASEB J, May 1, 1998; 12(7): 613 - 619.
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Q. Gu and R. L Moss
Novel mechanism for non-genomic action of 17{beta}-oestradiol on kainate-induced currents in isolated rat CA1 hippocampal neurones
J. Physiol., February 1, 1998; 506(3): 745 - 754.
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J. Tassignon and F. Haeseleer Abraham Borkowski
Natural Antiestrogen Receptor Autoantibodies in Man with Estrogenic Activity in Mammary Carcinoma Cell Culture: Study of their Mechanism of Action; Evidence for Involvement of Estrogen-Like Epitopes
J. Clin. Endocrinol. Metab., October 1, 1997; 82(10): 3464 - 3470.
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A. K. Morey, A. Pedram, M. Razandi, B. A. Prins, R.-M. Hu, E. Biesiada, and E. R. Levin
Estrogen and Progesterone Inhibit Vascular Smooth Muscle Proliferation
Endocrinology, August 1, 1997; 138(8): 3330 - 3339.
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V. Le Mellay, B. Grosse, and M. Lieberherr
Phospholipase C beta  and Membrane Action of Calcitriol and Estradiol
J. Biol. Chem., May 2, 1997; 272(18): 11902 - 11907.
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A. M. Nakhla, N. A. Romas, and W. Rosner
Estradiol Activates the Prostate Androgen Receptor and Prostate-specific Antigen Secretion through the Intermediacy of Sex Hormone-binding Globulin
J. Biol. Chem., March 14, 1997; 272(11): 6838 - 6841.
[Abstract] [Full Text] [PDF]

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C. Darimont, R. Delansorne, J. Paris, G. Ailhaud, and R. Negrel
Influence of Estrogenic Status on the Lipolytic Activity of Parametrial Adipose Tissue in Vivo: an in Situ Microdialysis Study
Endocrinology, March 1, 1997; 138(3): 1092 - 1096.
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K. Hisamoto, M. Ohmichi, H. Kurachi, J. Hayakawa, Y. Kanda, Y. Nishio, K. Adachi, K. Tasaka, E. Miyoshi, N. Fujiwara, et al.
Estrogen Induces the Akt-dependent Activation of Endothelial Nitric-oxide Synthase in Vascular Endothelial Cells
J. Biol. Chem., January 26, 2001; 276(5): 3459 - 3467.
[Abstract] [Full Text] [PDF]

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S. Kahlert, S. Nuedling, M. van Eickels, H. Vetter, R. Meyer, and C. Grohe
Estrogen Receptor alpha Rapidly Activates the IGF-1 Receptor Pathway
J. Biol. Chem., June 9, 2000; 275(24): 18447 - 18453.
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