TitleSolubilization and pharmacological characterization of a glucocorticoid membrane receptor from an amphibian brain
Publication TypeJournal Article
Year of Publication1998
AuthorsEvans, SJ, Moore, FL, Murray, TF
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume67
Pagination1-8
Type of ArticleJournal Article
ISSN0960-0760
Abstract

Physiological functions of steroid hormones involve activation of intracellular receptors as well as poorly understood membrane receptors. We report the pharmacological characterization of a solubilized corticosterone receptor from neuronal membranes. This receptor previously was shown to localize with plasma membrane subcellular fractions and to be involved in the modulation of courtship behaviors in the roughskin newt (Taricha granulosa). We describe procedures with non-ionic detergents that solubilize the receptor and maintain high affinity [H-3]corticosterone binding. The pharmacology of the solubilized corticosterone receptor resembles that of the membrane receptor with high affinity for [H-3]corticosterone and an identical rank-order potency for other steroid ligands (corticosterone>cortisol>aldosterone>dexamethasone). Unlike binding in membrane preparations, [H-3]corticosterone binding to the solubilized receptor is insensitive to negative modulation by guanyl nucleotides and only modestly sensitive to the presence of Mg2+. We also identified two ligands that exhibit high affinity binding to the solubilized receptor and have the potential to be used in an affinity purification scheme. They are corticosterone-3-carboxymethyloxime (CORT-3-CMO), which may be covalently attached to a Sepharose resin, and a derivitized azide form of CORT-3-CMO which can be covalently coupled to the solubilized receptor itself. The stability of the solubilized [H-3]corticosterone receptor in the detergent system will facilitate further purification and molecular characterization. (C) 1998 Elsevier Science Ltd. All rights reserved.

URL<Go to ISI>://WOS:000076064900001
DOI10.1016/s0960-0760(98)00070-3