title: Characterization and solubilization of novel aldosterone-binding proteins in porcine liver microsomes.
abstract: Using the radioligand [1,2,6,7-3H]aldosterone ([3H]aldosterone), specific binding sites for aldosterone were identified and characterized in microsomal preparations from porcine liver. The maximum binding capacity is approximately 700 fmol x mg-1 microsomal protein. The reversible binding of [3H]aldosterone was saturable and Scatchard analysis revealed two apparent dissociation constants (Kd), Kd1 < or = 11 nM and Kd2 = 118 nM. Binding was optimal at pH 7.2, thermolabile, and was reduced by more than 70% when membrane vesicles were pretreated with trypsin. Binding was selective for aldosterone with cortisol being a weak agonist at 1000-fold higher concentrations only. Among those detergents tested to optimize conditions for solubilization, n-octylglucoside (75 mM) was most favorable and solubilized 25% of the radioligand-binding protein complex in the undissociated form. These binding sites have unique pharmacological properties, which are similar to those found for aldosterone membrane binding in human lymphocytes and pig kidney, and for rapid aldosterone effects on sodium-proton exchange.
authors.affiliation: Medizinische Klinik, Klinikum Innenstadt, University of Munich, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Meyer
authors.forename: M
authors.initials: M
authors.lastname: Christ
authors.forename: M
authors.initials: M
authors.lastname: Wehling
mesh_terms: D000450:Aldosterone / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000032:analysis* / Q000737:chemistry; D006854:Hydrocortisone / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008862:Microsomes, Liver / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D018161:Receptors, Mineralocorticoid / Q000378:metabolism; D012995:Solubility; D013552:Swine; D014357:Trypsin / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D018161:Receptors, Mineralocorticoid; D000450:Aldosterone; D014357:Trypsin; D006854:Hydrocortisone