Characterization of the molybdate-stabilized glucocorticoid receptor from rat thymus.
The untransformed glucocorticoid receptor of rat thymus cytosol was characterized in the form of its complex with [1,2,4-3H]triamcinolone acetonide by ion-exchange chromatography and by gel filtration and sucrose-density-gradient ultracentrifugation at different ionic strengths. Molybdate (10 mM) was present throughout all experimental procedures and prevented receptor inactivation and degradation as well as transformation. At low ionic strength the molybdate-stabilized steroid-receptor complex was detected as a single highly asymmetric entity with a Stokes radius of 5.85 nm, a sedimentation coefficient of 9.6 S and an apparent molecular weight of 236 000. This form was converted into a smaller, even more asymmetric, form in increasing proportion as the ionic strength was increased. In the presence of 0.4 M-KCl, the smaller form had a Stokes radius of 4.95 nm, a sedimentation coefficient of 4.6 S and an apparent molecular weight of 95 500. It is concluded that the glucocorticoid-receptor complex exists at low ionic strengths as a homodimer or as a heterodimer in which only one subunit possesses a steroid-binding site, and that the process of dissociation into subunits brought about by increasing the ionic strength is a process distinct from, but possibly preceding, the transformation phenomenon responsible for conferring DNA-binding properties on the complex.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008297:Male; D008982:Molybdenum / Q000494:pharmacology*; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D013950:Thymus Gland / Q000378:metabolism*; D014222:Triamcinolone Acetonide / Q000378:metabolism
D046911:Macromolecular Substances; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D011189:Potassium Chloride; D008982:Molybdenum; C024687:sodium molybdate(VI); D014222:Triamcinolone Acetonide