The use of a hydroxylapatite-filter steroid receptor assay method in the study of the modulation of androgen receptor interaction.
Receptors for androgen, estrogen, and glucocorticoid can be assayed by hydroxylapatite adsorption of the radioactive steroid-receptor complex and washing of the adducts on membrane filters mounted on a multiple filter holder. The method is economical, very rapid and sensitive. This new receptor assay method was used to study the modulation of androgen receptor of rat ventral prostate by metal ions, thiols, and ligand structure. The interaction of androgen with the naked receptor is inhibited by 10 microM ZnCl2, CdSO4, or CuSO4 but this inhibition is competed by androgen and is reversed by DTT. The androgen-receptor complex is less sensitive to divalent metal ions but Zn2+, at 3 mM, appears to alter the conformation of the receptor and promote the release of androgen. Certain phenanthrene derivatives exhibited striking structural specificities in their ability to compete with radioactive androgen for binding to the prostate receptor. The results suggest that the receptor has binding preference toward individual ring structure in the steroid.
D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D017886:Durapatite; D004963:Estrenes / Q000378:metabolism; D005374:Filtration / Q000379:methods; D006882:Hydroxyapatites; D008297:Male; D015741:Metribolone; D008954:Models, Biological; D009281:Naphthalenes / Q000494:pharmacology; D010616:Phenanthrenes / Q000494:pharmacology; D011467:Prostate / Q000378:metabolism; D051381:Rats; D011944:Receptors, Androgen / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D013329:Structure-Activity Relationship
D000873:Anthracenes; D002413:Cations, Divalent; D004963:Estrenes; D006882:Hydroxyapatites; D009281:Naphthalenes; D010616:Phenanthrenes; D011944:Receptors, Androgen; D011987:Receptors, Steroid; C031721:naphthalene; D015741:Metribolone; C031181:phenanthrene; D017886:Durapatite; C034020:anthracene; D004229:Dithiothreitol