title: Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy.
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.
authors.affiliation: Biochemistry and Molecular Biology Laboratory, Centre of Advanced Study, Department of Zoology, Banaras Hindu University, Varanasi, UP, 221 005, India.
authors.forename: M K
authors.initials: MK
authors.lastname: Thakur
authors.affiliation: Biochemistry and Molecular Biology Laboratory, Centre of Advanced Study, Department of Zoology, Banaras Hindu University, Varanasi, UP, 221 005, India. vijayparamanik@gmail.com.|Department of Zoology, Indira Gandhi National Tribal University, Kapil Dhara Road, Mekal Sadan, Amarkantak, Anuppur, MP, 484 887, India. vijayparamanik@gmail.com.
authors.forename: V
authors.initials: V
authors.lastname: Paramanik
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism*; D047629:Estrogen Receptor beta / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D011485:Protein Binding; D016622:Silver Staining; D057188:Workflow
chemical_list: D047629:Estrogen Receptor beta; D004967:Estrogens; D004958:Estradiol
keywords: Cloning; Estradiol; Estrogen receptors; Nuclear magnetic resonance spectroscopy; SDS-polyacrylamide gel electrophoresis