Binding of aldosterone to cytoplasmic and nuclear receptors of the urinary bladder epithelium of Bufo marinus.

Binding of aldosterone to cytoplasmic and nuclear sites in urinary bladder epithelia of Bufo marinus (Dominican variant) is saturable and dependent upon steroid concentration. Scatchard analysis of specific cytoplasmic binding yielded a maximal binding capacity (N) of 14.5 X 10(-14) mol/mg protein and an apparent equilibrium dissociation constant (Kd) of 1.4 X 10(-8) M. Since Scatchard analysis of specific nuclear binding was complex, this binding was resolved by a computer-generated cirve-fitting technique which analyzed total aldosterone bound. Nuclear binding was resolved into three sites: a nonsaturable site that was linearly dependent upon aldosterone concentration, and two saturable sites (types I and II). Type I sites had relatively low capacity for aldosterone (N = 31 +/- 1 X 10(-14) mol/mg DNA) and high affinity (Kd = 2.5 +/- 0.5 X 10(-9 M); tffininty (Kd = 8.6 +/- 1.7 X 10(-7) M). Competition for [3H]aldosterone binding by dexamethasone, corticosterone, cortisol, progesterone, testosterone, and 17 beta-estradiol demonstrated that type I nuclear sites have higher affinity for aldosterone than for other steroids. The findings are consistent with the inference that the type I site is the mineralocorticoid receptor.