The androgen receptor has been purified from rat ventral prostate cytosol by a combination of differential DNA-Sepharose 4B chromatography and testosterone 17β-hemisuccinyl-3,3′-diaminodipropylamine-Sepharose 4B affinity chromatography. Approximately 8 µg of protein was obtained from 38 g of rat ventral prostate, with a yield of 24%. The receptor was purified approximately 120000-fold. Silver nitrate staining of a sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel revealed a major polypeptide band migrating at 86 000 daltons. Affinity labeling of a partially purified receptor preparation with either 17-hydroxy-17α-[3H]methyl-4,9,11-estratrien-3-one or 17β-hydroxy-[1,2,4,5,6,7,16,17-3H8]-5α-androstan-3-one 17-(2-bromoacetate) produced a major band of radioactivity migrating at 86 000 daltons on a NaDodSO4 gel. Under nondenaturing conditions, a Mr of 85 000 was determined by gel filtration (42 Å) and sucrose gradient sedimentation analysis (4.5 S). The purified receptor had an isoelectric point of 6.3. [3H]-4,5α-Dihydrotestosterone, bound to the purified receptor, was displaced with 4,5α-dihydrotestosterone > testosterone ≫ progesterone > 5α-androstane-3α,17β-diol > 17β-estradiol > Cortisol. A number of physicochemical properties of the purified receptor were similar to those of the receptor in crude cytosol.
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