Expression, purification, and characterization of multiple, multifunctional human glucocorticoid receptor proteins

Protein Expr Purif. 2008 Nov;62(1):29-35. doi: 10.1016/j.pep.2008.07.008. Epub 2008 Jul 30.

Abstract

The glucocorticoid receptor (GR) is a nuclear receptor protein that plays a central role in glucose homeostasis, the stress response, control of the hypothalamic-pituitary-adrenal axis, and immuno-inflammatory processes via binding of the natural steroid, cortisol. GR is a well-validated drug target and continues to be an important target for new drug discovery efforts. Here, we describe a basic and simple method for Escherichia coli expression and purification of a variety of human GR proteins that contain all three of the functional domains of the protein: the activation function-1 domain, the DNA-binding domain, and the ligand-binding domain. We present characterization data to show that these purified, multifunctional GR proteins are active for ligand, coactivator, and DNA-binding. The work presented here should serve as a reference for future mechanistic, structural and drug discovery efforts that require purified, full or near full length, GR protein.

MeSH terms

  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Humans
  • Ligands
  • Receptors, Glucocorticoid / chemistry*
  • Receptors, Glucocorticoid / genetics
  • Receptors, Glucocorticoid / isolation & purification*

Substances

  • Ligands
  • Receptors, Glucocorticoid