Novel and simple two-step purification of a full-length rat glucocorticoid-receptor expressed in a baculovirus system

We purified the activated recombinant glucicorticoid receptor (GR) overexpr essed in insect cells by sequential chromatographies using Mono Q and Mono S columns. This procedure was based upon a new finding that the activated G R binds both to a Mono Q column and to a Mono S column at the same pH (pH 8 .4). The entire chromatographies took about 3 h and GR represented 97% of t he purified protein sample. The purified GR was able to bind specifically t o a DNA fragment containing the glucocorticoid response element. This purif ication protocol will be applicable to the purification of native GR, point -mutated recombinant GR and other nuclear receptors. (C) 2001 Elsevier Scie nce B.V. All rights reserved.