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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Acknowledgements
Financial support from UGC-CAS and Department of Biotechnology, Government of India, to M.K.T. is highly acknowledged.
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Thakur, M.K., Paramanik, V. (2016). Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy. In: Eyster, K.M. (eds) Estrogen Receptors. Methods in Molecular Biology, vol 1366. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3127-9_18
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DOI: https://doi.org/10.1007/978-1-4939-3127-9_18
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3126-2
Online ISBN: 978-1-4939-3127-9
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