Advanced

Backbone 1H, 13C, and 15N resonance assignments of the ligand binding domain of the human wildtype glucocorticoid receptor and the F602S mutant variant

Köhler, Christian; Carlström, Göran LU ; Tångefjord, Stefan; Papavoine, Tineke; Lepistö, Matti; Edman, Karl and Akke, Mikael LU (2018) In Biomolecular NMR Assignments 12(2). p.263-268
Abstract

The glucocorticoid receptor (GR) is a nuclear hormone receptor that regulates key genes controlling development, metabolism, and the immune response. GR agonists are efficacious for treatment of inflammatory, allergic, and immunological disorders. Steroid hormone binding to the ligand-binding domain (LBD) of GR is known to change the structural and dynamical properties of the receptor, which in turn control its interactions with DNA and various co-regulators and drive the pharmacological response. Previous biophysical studies of the GR LBD have required the use of mutant forms to overcome issues with limited protein stability and high aggregation propensity. However, these mutant variants are known to also influence the functional... (More)

The glucocorticoid receptor (GR) is a nuclear hormone receptor that regulates key genes controlling development, metabolism, and the immune response. GR agonists are efficacious for treatment of inflammatory, allergic, and immunological disorders. Steroid hormone binding to the ligand-binding domain (LBD) of GR is known to change the structural and dynamical properties of the receptor, which in turn control its interactions with DNA and various co-regulators and drive the pharmacological response. Previous biophysical studies of the GR LBD have required the use of mutant forms to overcome issues with limited protein stability and high aggregation propensity. However, these mutant variants are known to also influence the functional response of the receptor. Here we report a successful protocol for protein expression, purification, and NMR characterization of the wildtype human GR LBD. We achieved chemical shift assignments for 90% of the LBD backbone resonances, with 216 out of 240 non-proline residues assigned in the 1H–15N TROSY spectrum. These advancements form the basis for future investigations of allosteric effects in GR signaling.

(Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Allostery, Glucocorticoid receptor, Ligand binding, Nuclear receptors
in
Biomolecular NMR Assignments
volume
12
issue
2
pages
263 - 268
publisher
Springer
external identifiers
  • scopus:85045453713
ISSN
1874-2718
DOI
10.1007/s12104-018-9820-9
language
English
LU publication?
yes
id
0a3382dd-7d0e-4006-950f-3bc1d86a55ca
date added to LUP
2018-04-23 14:31:02
date last changed
2019-03-19 03:53:29
@article{0a3382dd-7d0e-4006-950f-3bc1d86a55ca,
  abstract     = {<p>The glucocorticoid receptor (GR) is a nuclear hormone receptor that regulates key genes controlling development, metabolism, and the immune response. GR agonists are efficacious for treatment of inflammatory, allergic, and immunological disorders. Steroid hormone binding to the ligand-binding domain (LBD) of GR is known to change the structural and dynamical properties of the receptor, which in turn control its interactions with DNA and various co-regulators and drive the pharmacological response. Previous biophysical studies of the GR LBD have required the use of mutant forms to overcome issues with limited protein stability and high aggregation propensity. However, these mutant variants are known to also influence the functional response of the receptor. Here we report a successful protocol for protein expression, purification, and NMR characterization of the wildtype human GR LBD. We achieved chemical shift assignments for 90% of the LBD backbone resonances, with 216 out of 240 non-proline residues assigned in the <sup>1</sup>H–<sup>15</sup>N TROSY spectrum. These advancements form the basis for future investigations of allosteric effects in GR signaling.</p>},
  author       = {Köhler, Christian and Carlström, Göran and Tångefjord, Stefan and Papavoine, Tineke and Lepistö, Matti and Edman, Karl and Akke, Mikael},
  issn         = {1874-2718},
  keyword      = {Allostery,Glucocorticoid receptor,Ligand binding,Nuclear receptors},
  language     = {eng},
  month        = {04},
  number       = {2},
  pages        = {263--268},
  publisher    = {Springer},
  series       = {Biomolecular NMR Assignments},
  title        = {Backbone <sup>1</sup>H, <sup>13</sup>C, and <sup>15</sup>N resonance assignments of the ligand binding domain of the human wildtype glucocorticoid receptor and the F602S mutant variant},
  url          = {http://dx.doi.org/10.1007/s12104-018-9820-9},
  volume       = {12},
  year         = {2018},
}