help button home button Endocrine Society Molecular Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Molecular Endocrinology, doi:10.1210/me.2002-0250
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
16/12/2706    most recent
Author Manuscript (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow NURSA Molecule Pages Link
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tamrazi, A.
Right arrow Articles by Katzenellenbogen, J. A
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tamrazi, A.
Right arrow Articles by Katzenellenbogen, J. A
Molecular Endocrinology 16 (12): 2706-2719
Copyright © 2002 by The Endocrine Society

Estrogen Receptor Dimerization: Ligand Binding Regulates Dimer Affinity and DimerDissociation Rate

Anobel Tamrazi, Kathryn E. Carlson, Jonathan R. Daniels, Kyle M. Hurth and John A Katzenellenbogen

Department of Chemistry, University of Illinois, Urbana, Illinois 61801

Address all correspondence and requests for reprints to: John A. Katzenellenbogen, Department of Chemistry, University of Illinois, 600 South Mathews Avenue, Urbana, Illinois 61801. E-mail: jkatzene{at}uiuc.edu.

Nuclear receptors form strong dimers that are essential for their function as transcription factors, and it is thought that ligand binding can affect dimer stability. In this report, we describe convenient fluorescence resonance energy transfer (FRET)-based methods for measuring the thermodynamic and kinetic stability of dimers of the estrogen receptor-{alpha} ligand-binding domain (ER{alpha}-LBD). We have developed receptors that are chemically labeled with a single fluorophore in a site-specific manner. These fluorophore-labeled ERs are functional and can be used to measure directly the affinity and stability of ER{alpha}-LBD dimers. Our results indicate that unliganded ER{alpha}-LBDs exist as very stable dimers and that the dissociation rate of these dimers is slow (t1/2&equals;39 ± 3 min at 28 C) and is further slowed (<=7-fold) by the addition of various ligands. Estrogen antagonists provide greater kinetic stabilization of the ER dimers than agonists. In addition, coactivator peptides containing the LXXLL motif selectively stabilize agonist-bound ER{alpha}-LBD dimers. These fluorescence-based assays for measuring the kinetic and thermodynamic stability of ER dimers provide a functional in vitro method for assessing the agonist or antagonist character of novel ligands.

NURSA Molecule Pages Link:

Nuclear Receptors:   ERα
Coregulators:   SRC-1
Ligands:   17β-Estradiol  |  Diethylstilbestrol  |  4-Hydroxytamoxifen  |  Raloxifene



This article has been cited by other articles:


Home page
 Mol. Endocrinol.Home page
M. T. Sonoda, L. Martinez, P. Webb, M. S. Skaf, and I. Polikarpov
Ligand Dissociation from Estrogen Receptor Is Mediated by Receptor Dimerization: Evidence from Molecular Dynamics Simulations
Mol. Endocrinol., July 1, 2008; 22(7): 1565 - 1578.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
R. Paulmurugan, A. Tamrazi, J. A. Katzenellenbogen, B. S. Katzenellenbogen, and S. S. Gambhir
A Human Estrogen Receptor (ER){alpha} Mutation with Differential Responsiveness to Nonsteroidal Ligands: Novel Approaches for Studying Mechanism of ER Action
Mol. Endocrinol., July 1, 2008; 22(7): 1552 - 1564.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. Y. Dai, M. J. Chalmers, J. Bruning, K. S. Bramlett, H. E. Osborne, C. Montrose-Rafizadeh, R. J. Barr, Y. Wang, M. Wang, T. P. Burris, et al.
Prediction of the tissue-specificity of selective estrogen receptor modulators by using a single biochemical method
PNAS, May 20, 2008; 105(20): 7171 - 7176.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
S. R. Hammes and E. R. Levin
Extranuclear Steroid Receptors: Nature and Actions
Endocr. Rev., December 1, 2007; 28(7): 726 - 741.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. E. Ramsey, M. A. Daugherty, and R. J. Kelm Jr.
Hydrodynamic Studies on the Quaternary Structure of Recombinant Mouse Purbeta
J. Biol. Chem., January 19, 2007; 282(3): 1552 - 1560.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
A. Padron, L. Li, E. M. Kofoed, and F. Schaufele
Ligand-Selective Interdomain Conformations of Estrogen Receptor-{alpha}
Mol. Endocrinol., January 1, 2007; 21(1): 49 - 61.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
S. Khan, R. Barhoumi, R. Burghardt, S. Liu, K. Kim, and S. Safe
Molecular Mechanism of Inhibitory Aryl Hydrocarbon Receptor--Estrogen Receptor/Sp1 Cross Talk in Breast Cancer Cells
Mol. Endocrinol., September 1, 2006; 20(9): 2199 - 2214.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
M. Abdelrahim, E. Ariazi, K. Kim, S. Khan, R. Barhoumi, R. Burghardt, S. Liu, D. Hill, R. Finnell, B. Wlodarczyk, et al.
3-Methylcholanthrene and Other Aryl Hydrocarbon Receptor Agonists Directly Activate Estrogen Receptor {alpha}
Cancer Res., February 15, 2006; 66(4): 2459 - 2467.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
A. Tamrazi, K. E. Carlson, A. L. Rodriguez, and J. A. Katzenellenbogen
Coactivator Proteins as Determinants of Estrogen Receptor Structure and Function: Spectroscopic Evidence for a Novel Coactivator-Stabilized Receptor Conformation
Mol. Endocrinol., June 1, 2005; 19(6): 1516 - 1528.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
K. Kim, R. Barhoumi, R. Burghardt, and S. Safe
Analysis of Estrogen Receptor {alpha}-Sp1 Interactions in Breast Cancer Cells by Fluorescence Resonance Energy Transfer
Mol. Endocrinol., April 1, 2005; 19(4): 843 - 854.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
S. H. Kim, A. Tamrazi, K. E. Carlson, and J. A. Katzenellenbogen
A Proteomic Microarray Approach for Exploring Ligand-initiated Nuclear Hormone Receptor Pharmacology, Receptor Selectivity, and Heterodimer Functionality
Mol. Cell. Proteomics, March 1, 2005; 4(3): 267 - 277.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
M. Razandi, A. Pedram, I. Merchenthaler, G. L. Greene, and E. R. Levin
Plasma Membrane Estrogen Receptors Exist and Functions as Dimers
Mol. Endocrinol., December 1, 2004; 18(12): 2854 - 2865.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
C. L. Smith and B. W. O'Malley
Coregulator Function: A Key to Understanding Tissue Specificity of Selective Receptor Modulators
Endocr. Rev., February 1, 2004; 25(1): 45 - 71.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
A. Tamrazi, K. E. Carlson, and J. A. Katzenellenbogen
Molecular Sensors of Estrogen Receptor Conformations and Dynamics
Mol. Endocrinol., December 1, 2003; 17(12): 2593 - 2602.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals
Copyright © 2002 by The Endocrine Society