title: NMR analysis reveals 17β-estradiol induced conformational change in ERβ ligand binding domain expressed in E. coli.
abstract: Nuclear magnetic resonance (NMR) spectroscopy is a useful biophysical technique to study the ligand-protein interaction. In this report, we have used bacterially produced ERβ and its domains for studying the functional analysis of ligand-protein interaction. Briefly, ERβ and its transactivation domain (TAD) and ligand binding domain (LBD) were subcloned and overexpressed using a prokaryotic expression system. The recombinant proteins were purified using Ni(+2)-IDA affinity chromatography and analyzed by NMR. Purified ERβ and TAD show similar conformation in the absence or presence of 17β-estradiol. However, LBD shows altered conformation in the presence of 17β-estradiol. These findings suggest that ERβ produced in bacteria exhibits a conformation such that its LBD remains masked and consequently it binds less to 17β-estradiol. Such study may help to develop the therapeutic approaches for controlling the estradiol-mediated gene expression in hormone dependent diseases.
authors.affiliation: Department of Zoology, Biochemistry and Molecular Biology Laboratory, Centre of Advanced Study, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Vijay
authors.initials: V
authors.lastname: Paramanik
authors.forename: M K
authors.initials: MK
authors.lastname: Thakur
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology*; D047629:Estrogen Receptor beta / Q000737:chemistry* / Q000378:metabolism*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D047629:Estrogen Receptor beta; D008024:Ligands; D011994:Recombinant Proteins; D004958:Estradiol