Aliquots of @entity35 liver cytosol glucocorticoid-receptor complexes (GRc) were transformed by an incubation with 8-10 mM @entity855 at 0 degrees C and were compared with those transformed by an exposure to 23 degrees C. The extent of receptor transformation was measured by chromatography of the samples over columns of @entity8851 -Sephacel. The @entity855 -transformed complexes, like those which were heat-transformed, exhibited lower affinity for the positively charged ion-exchange resin and were eluted with 0.12 M KCl (peak-I): the nontransformed complexes appeared to possess higher affinity and required 0.21 M @entity4338 (peak II) for their elution. As expected, the receptor in the peak-I exhibited the DNA-cellulose binding capacity and sedimented as 4S in @entity969 gradients. Peak II contained an 8-9S glucocorticoid receptor (GR) form that showed reduced affinity for DNA-cellulose. Presence of @entity55337 (5 mM) prevented both heat and @entity855 transformation of the GRc resulting in the elution of the complexes in the region of nontransformed receptors. When parallel experiments were performed, binding of the cytosol GRc to @entity35 liver nuclei or DNA-cellulose was seen to increase 10-15 fold upon transformation by heat or @entity855 : @entity45746 treatment blocked this process completely. The transformed and nontransformed GRc were also differentially fractionated by @entity4859 : tungstate-treated (nontransformed) receptor required higher salt concentration and was precipitated at 55% saturation. In addition, the GRc could be extracted from DNA-cellulose by an incubation of the affinity resin with @entity55337 resulting in approximately 500-fold purification of the receptor with a 30% yield. These studies show that the nontransformed, and the heat-, salt-, and @entity855 -transformed GRc from the @entity35 liver cytosol can be separated chromatographically, and that the use of tungstate facilitates the resolution of these different receptor forms. In addition, extraction of the receptor from DNA-cellulose by tungstate provides another new and efficient method of partial receptor purification.

Chromatographic separation of nontransformed and transformed glucocorticoid-receptor complexes from XXXX liver cytosol. Use of tungstate in the purification, inactivation, and resolution of receptor forms.

@entity55337 :: ('MESH:C025399', 'Chemical') :: ['sodium tungstate'] @entity4859 :: ('CHEBI:62946', 'Chemical') :: ['(NH4)2SO4'] @entity969 :: ('MESH:D013395', 'Chemical') :: ['sucrose'] @entity8851 :: ('MESH:C007369', 'Chemical') :: ['DEAE'] @entity45746 :: ('MESH:C045951', 'Chemical') :: ['tungstate'] @entity35 :: ('10116', 'Species') :: ['rat'] @entity4338 :: ('CHEBI:32588', 'Chemical') :: ['KCl'] @entity855 :: ('MESH:D000255', 'Chemical') :: ['ATP']
@entity35 :: (10116,Species) :: ['rat']
