Transformation of the [3H]aldosterone-type I receptor complex to a DNA-binding species.
For most steroid receptor complexes, the transformation to a DNA-binding species can be achieved readily in vitro by incubation at elevated temperatures and/or salt concentrations. Although the aldosterone-Type I receptor complex forms a clear exception to this generalization, a marked increase in its transformation can be achieved by incubation with the chaotropic anion, thiocyanate. Time and concentration-response analyses with brain cytosol revealed that over 40% of the complexes were retained in DNA-cellulose assays after a 15 min pre-incubation at 0 degree C with 100 mM thiocyanate. As expected, molybdate prevented this transformation; however, in contrast to results with heat- and/or salt-induced transformation of other steroid receptors, the molybdate effect was only partially removed by gel filtering the cytosol prior to thiocyanate addition. Thiocyanate-induced transformation should prove useful in the biochemical characterization and purification of non-transformed and transformed aldosterone-Type I receptor complexes.
Luttge
W G
WG
Department of Neuroscience, University of Florida College of Medicine, Gainesville 32610.
Emadian
S M
SM
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002482:Cellulose / Q000031:analogs & derivatives / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000031:analogs & derivatives / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D051379:Mice; D008982:Molybdenum / Q000494:pharmacology; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000378:metabolism*; D018161:Receptors, Mineralocorticoid; D013696:Temperature; D013861:Thiocyanates / Q000494:pharmacology*
C033190:DNA-cellulose; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; D013861:Thiocyanates; C044659:molybdate; D008982:Molybdenum; D002482:Cellulose; D004247:DNA; C031760:thiocyanate