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Updated: Mar 30, 2026

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
Published on: December 19, 2018
Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy
M K Thakur1, V Paramanik2,3
1Biochemistry and Molecular Biology Laboratory, Centre of Advanced Study, Department of Zoology, Banaras Hindu University, Varanasi, UP, 221 005, India.
Abstract:
Following binding to estradiol, estrogen receptors (ER) α and ERβ recruit a number of interacting proteins and mediate a plethora of functions. The binding of estrogen with the receptors shows changes in the resonance structure and movement of protons. We cloned ERβ and its trans-activation domain (TAD) and ligand-binding domain (LBD), expressed them in prokaryotic expression vectors, purified them, and studied their interaction with estradiol. In this chapter, a detailed method of preparation of recombinant proteins, SDS-PAGE, silver staining, and NMR are described. Such methods are useful to check the biological activity of bacterially expressed proteins and are applicable to basic and applied research.
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