Binding of aldosterone by epidermal cytosol in the tail of bullfrog larvae.
Characteristics of aldosterone binding by epidermal cytosol in the tail of the larval bullfrog were investigated. Temperature-dependent binding experiments showed that the specific binding of [3H]aldosterone to the tail cytosol was thermolabile. A 3-hr incubation at 0 degrees was the optimum assay condition required for reaching equilibrium. Separation of bound and free hormone was performed using a hydroxylapatite method. Specific binding of aldosterone was observed in the tail epidermis but not in the tail mesenchyme. Saturation analysis revealed that specific binding of [3H]aldosterone to the epidermal cytosol reached maximum between 20 and 40 nM. Scatchard analysis for the cytosol of the tail epidermis yielded a straight line with a dissociation constant (Kd) of 7.0 nM and the maximum number of binding sites (Bmax) of 56.4 fmol/mg protein. Sucrose density gradient analysis of crude cytosol revealed a specific peak of radioactivity in the 8-9 S area. Steroid-binding specificity revealed a significant displacement of the [3H]aldosterone by radioinert aldosterone and, to a lesser extent, by cortisol, whereas 17 beta-estradiol and testosterone competed very poorly. However, corticosterone and dexamethasone, glucocorticoids in mammals, were better competitors of the aldosterone binding, whereas ZK91587, a selective synthetic antagonist for mineralocorticoid receptor in mammals, scarcely inhibited the aldosterone binding. These results suggest that the aldosterone-binding site may also be a corticosterone receptor and that there may be no other receptors specific for aldosterone at least in the tail epidermis.
Yamamoto
K
K
Department of Biology, School of Education, Waseda University, Tokyo, Japan.
Kikuyama
S
S
D000450:Aldosterone / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003600:Cytosol / Q000378:metabolism*; D004817:Epidermis / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D007814:Larva; D008970:Molecular Weight; D011892:Rana catesbeiana; D018161:Receptors, Mineralocorticoid / Q000032:analysis / Q000378:metabolism; D013623:Tail; D013696:Temperature
D018161:Receptors, Mineralocorticoid; D000450:Aldosterone