Purification and characterization of the androgen receptor from rat ventral prostate

Ching H. Chang, David R. Rowley, Donald J. Tindall

Research output: Contribution to journalArticle

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Abstract

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,l l-estratrien-3-one or 17β-hydroxy-[1,2,4,5,6,7,16,17-3H 8]-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 85000 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.

Original languageEnglish (US)
Pages (from-to)6170-6175
Number of pages6
JournalBiochemistry
Volume22
Issue number26
StatePublished - 1983
Externally publishedYes

Fingerprint

Dihydrotestosterone
Androgen Receptors
Cytosol
Sepharose
Androstane-3,17-diol
Purification
Testosterone
Rats
Prostate
Gels
Affinity chromatography
Silver Nitrate
Agarose Chromatography
Silver Staining
Radioactivity
Isoelectric Point
Chromatography
Affinity Chromatography
Sedimentation
Sodium Dodecyl Sulfate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Chang, C. H., Rowley, D. R., & Tindall, D. J. (1983). Purification and characterization of the androgen receptor from rat ventral prostate. Biochemistry, 22(26), 6170-6175.

Purification and characterization of the androgen receptor from rat ventral prostate. / Chang, Ching H.; Rowley, David R.; Tindall, Donald J.

In: Biochemistry, Vol. 22, No. 26, 1983, p. 6170-6175.

Research output: Contribution to journalArticle

Chang, CH, Rowley, DR & Tindall, DJ 1983, 'Purification and characterization of the androgen receptor from rat ventral prostate', Biochemistry, vol. 22, no. 26, pp. 6170-6175.
Chang, Ching H. ; Rowley, David R. ; Tindall, Donald J. / Purification and characterization of the androgen receptor from rat ventral prostate. In: Biochemistry. 1983 ; Vol. 22, No. 26. pp. 6170-6175.
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