Chapter title |
Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy
|
---|---|
Chapter number | 18 |
Book title |
Estrogen Receptors
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3127-9_18 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3126-2, 978-1-4939-3127-9
|
Authors |
M. K. Thakur, V. Paramanik, Thakur, M. K., Paramanik, V. |
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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