Physicochemical characterization of the androgen receptor from hyperplastic human prostate

L. R. Murthy, C. H. Chang, D. R. Rowley, P. T. Scardino, D. J. Tindall

Research output: Contribution to journalArticle

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Abstract

In this manuscript we have characterized the androgen receptor from human benign prostatic hypertrophied (BPH) tissue. BPH tissue was obtained fresh from surgery, ground, and frozen in liquid nitrogen. Tissue was homogenized in 10 mM Tes buffer (pH 7.4 at 25°C) containing 0.25 M sucrose, 20 mM Na2MoO4, 12 mM monothioglycerol, and 1.5 mM EDTA. Cytosol was labeled with (3H)-R1881 (0.5-10 nM) ± a 100-fold excess of R1881 and a 500-fold excess of triamcinolone acetonide to identify androgen receptor sites. An exchange assay was developed, whereby maximum binding was achieved by raising the temperature to 30°C for 15 min. A high-affinity binding protein was detected (Kd = 1.3 ± 0.8 nM) which had a binding capacity of 15 ± 7 fmol/mg protein. Binding was specific for androgens with the following Ki (nM) values: R1881 (0.09), dihydrotestosterone (6), testosterone (50), progesterone (92), estradiol (100), epitestosterone (860), and cortisol (> 1,000). Under high ionic conditions the sedimentation, coefficient of the receptor was 4.1 S. Higher S values were not observed in the presence of the following protein inhibitors: leupeptin (20 or 150 μM), iodoacetamide (10 mM), DFP (5 mM), PMSF (1 mM), bacitracin (0.1 mM), antipain (0.1 mM), aprotinin (1 TIU/ml. Gel filtration analysis revealed a Stokes radius of 6.5 nm. These data indicate a M(r) of 120,000 and a f/fo of 2.01. The receptor eluted from a chromatofocusing column at a pH of 6.8. The receptor bound to phosphocellulose and DNA cellulose after being heat-treated for 15 min at 30°C. These results suggest that BPH tissue contains an androgen receptor which is similar to receptors in other androgen-responsive tissues.

Original languageEnglish (US)
Pages (from-to)567-579
Number of pages13
JournalProstate
Volume5
Issue number6
DOIs
StatePublished - 1984
Externally publishedYes

Fingerprint

Metribolone
Prostate
Androgen Receptors
Androgens
Epitestosterone
Antipain
Isoflurophate
Bacitracin
Iodoacetamide
Triamcinolone Acetonide
Aprotinin
Dihydrotestosterone
Edetic Acid
Cytosol
Gel Chromatography
Progesterone
Sucrose
Hydrocortisone
Testosterone
Estradiol

ASJC Scopus subject areas

  • Urology

Cite this

Murthy, L. R., Chang, C. H., Rowley, D. R., Scardino, P. T., & Tindall, D. J. (1984). Physicochemical characterization of the androgen receptor from hyperplastic human prostate. Prostate, 5(6), 567-579. https://doi.org/10.1002/pros.2990050602

Physicochemical characterization of the androgen receptor from hyperplastic human prostate. / Murthy, L. R.; Chang, C. H.; Rowley, D. R.; Scardino, P. T.; Tindall, D. J.

In: Prostate, Vol. 5, No. 6, 1984, p. 567-579.

Research output: Contribution to journalArticle

Murthy, LR, Chang, CH, Rowley, DR, Scardino, PT & Tindall, DJ 1984, 'Physicochemical characterization of the androgen receptor from hyperplastic human prostate', Prostate, vol. 5, no. 6, pp. 567-579. https://doi.org/10.1002/pros.2990050602
Murthy, L. R. ; Chang, C. H. ; Rowley, D. R. ; Scardino, P. T. ; Tindall, D. J. / Physicochemical characterization of the androgen receptor from hyperplastic human prostate. In: Prostate. 1984 ; Vol. 5, No. 6. pp. 567-579.
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