title: Occurrence of glucocorticoid binding sites in solubilized microsomes from rat liver.
abstract: Recent studies suggested the presence of specific glucocorticoid binding sites on rat liver microsomal membranes. We report here a new solubilization procedure which allows the physicochemical characterization of the microsomal glucocorticoid binding sites. Solubilization was achieved with 2 mM CHAPS in the presence of 5 mM benzamidine. Binding of [3H]cortisol showed a high affinity (Kd = 5.1 x 10(-9) M) and a limited capacity (0.72 pmol/mg of protein). The binding activity was abolished by elevated temperature and pronase. Competition experiments revealed that natural glucocorticoids and progesterone were highly potent competitors whereas dexamethasone and triamcinolone acetonide did not compete. Chromatography on DEAE Trisacryl and heparin Ultrogel confirmed that the solubilized protein is different from corticosteroid binding globulin and the cytosolic glucocorticoid receptor. Treatment of microsomal fractions with phosphatidyl inositol phospholipase C promoted the release of specific binding activity suggesting a putative glycosylphosphatidyl anchor for this protein. This finding may have interesting implications concerning the mechanism of glucocorticoid hormone action.
authors.affiliation: Department of Medical Biochemistry, Nantes University Hospital, France.
authors.forename: F
authors.initials: F
authors.lastname: Guendouz
authors.forename: P
authors.initials: P
authors.lastname: Blanchardie
authors.forename: M
authors.initials: M
authors.lastname: Denis
authors.forename: J L
authors.initials: JL
authors.lastname: Orsonneau
authors.forename: P
authors.initials: P
authors.lastname: Lustenberger
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002458:Cell Fractionation; D002852:Chromatography, Ion Exchange; D006854:Hydrocortisone / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism* / Q000648:ultrastructure; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011965:Receptors, Glucocorticoid / Q000302:isolation & purification / Q000378:metabolism*; D010738:Type C Phospholipases / Q000494:pharmacology
chemical_list: D011480:Protease Inhibitors; D011965:Receptors, Glucocorticoid; D010738:Type C Phospholipases; D006854:Hydrocortisone