Purification and characterization of androgen receptor from steer seminal vesicle

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

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

The androgen receptor has been purified from steer seminal vesicle cytosol by a combination of differential DNA-Sepharose 4B chromatography and testosterone 17β-hemisuccinyl-3,3′-diaminodipropylamine-Sepharose 4B affinity chromatography. The procedure produced about 3 μg of receptor protein from 35 g of steer seminal vesicle, with a yield of 48%. The receptor protein, as a complex with unlabeled testosterone, was purified approximately 540 000-fold. A single band, migrating at 60 000 daltons, was observed following electrophoresis on a polyacrylamide gel containing sodium dodecyl sulfate (NaDodSO4). This was confirmed by affinity labeling of the partially purified receptor with both 17-hydroxy-17α-[3H]methyl-4,9,11-estratrien-3-one and 17β-hydroxy-[1,2,4,5,6,7,16,17-3H 8]-5α-androstan-3-one 17-(2-bromoacetate), which showed a peak of radioactivity migrating at 60 000 daltons by NaDodSO4 gel electrophoresis. The receptor had an estimated Stokes radius of 35 Å and a sedimentation coefficient of 3.8 S in the presence of 0.3 M NaCl. The calculated molecular weight and frictional ratio for the androgen binding activity were 57 000 and 1.42, respectively. Chromatofocusing of the purified receptor protein revealed an isoelectric point of 6.6. [3H]Methyltrienolone, bound to the purified receptor, was displaced with methyltrienolone > testosterone > 5α-dihydrotestosterone ≫ 3β-hydroxy-Δ5-androsten-17-one ≫ 5α-androstane-3α,17β-diol. The physicochemical properties of the purified receptor were similar to those of the receptor in crude cytosol. These results demonstrate that the androgen receptor from steer seminal vesicle was substantially purified without significant modification of its physicochemical or steroid binding properties.

Original languageEnglish (US)
Pages (from-to)4102-4109
Number of pages8
JournalBiochemistry
Volume21
Issue number17
StatePublished - 1982
Externally publishedYes

Fingerprint

Seminal Vesicles
Androgen Receptors
Metribolone
Purification
Testosterone
Electrophoresis
Cytosol
Sepharose
Androstane-3,17-diol
Affinity chromatography
Agarose Chromatography
Proteins
Dihydrotestosterone
Radioactivity
Isoelectric Point
Chromatography
Affinity Chromatography
Sedimentation
Sodium Dodecyl Sulfate
Labeling

ASJC Scopus subject areas

  • Biochemistry

Cite this

Chang, C. H., Rowley, D. R., Lobl, T. J., & Tindall, D. J. (1982). Purification and characterization of androgen receptor from steer seminal vesicle. Biochemistry, 21(17), 4102-4109.

Purification and characterization of androgen receptor from steer seminal vesicle. / Chang, Ching H.; Rowley, David R.; Lobl, Thomas J.; Tindall, Donald J.

In: Biochemistry, Vol. 21, No. 17, 1982, p. 4102-4109.

Research output: Contribution to journalArticle

Chang, CH, Rowley, DR, Lobl, TJ & Tindall, DJ 1982, 'Purification and characterization of androgen receptor from steer seminal vesicle', Biochemistry, vol. 21, no. 17, pp. 4102-4109.
Chang CH, Rowley DR, Lobl TJ, Tindall DJ. Purification and characterization of androgen receptor from steer seminal vesicle. Biochemistry. 1982;21(17):4102-4109.
Chang, Ching H. ; Rowley, David R. ; Lobl, Thomas J. ; Tindall, Donald J. / Purification and characterization of androgen receptor from steer seminal vesicle. In: Biochemistry. 1982 ; Vol. 21, No. 17. pp. 4102-4109.
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