Characterization of the cytosol androgen receptor of the human prostate

Dirk M. Wilbert, Jim Griffin III, Jean D. Wilson

Research output: Contribution to journalArticle

102 Citations (Scopus)

Abstract

Direct measurement of the binding of endogenous androgens to the androgen receptor of human tissues has not been possible because of contamination of tissue with traces of plasma proteins, such as testosterone-binding globulin (TeBG), that contain more androgen-binding capacity than does the receptor itself. Molybdate is known to stabilize the 8–9S forms of other steroid hormone receptors. We took advantage of this phenomenon tocharacterize the androgen receptor of hyperplastic prostates removed at surgery, using sucrose density gradient centrifugation in a vertical rotor. In 10 mM sodium molybdate, theandrogen receptor sediments as a distinct 9.2 ± 0.5S moiety, easily separable from TeBG. Unlike TeBG, the 9S receptor is not removed by absorption with Concanavalin A. [3H]Dihydrotestosterone (3H-labeled 17β-hydroxy-5α-androstan- 3-one) bindingto the 9S receptor is not competed for by excess triamcinoloneacetonide (9α-fluoro-llβ,16α,17α,21-tetrahydroxypregna- l,4-diene-3,20-dione cyclic 16,17-acetonide) or promegestone (17,21-dimethyl-19-non-pregna-4,9-diene-3,20- dione), which are known to bind to the progestin receptor. In contrast, [3H]methyltrienolone (3H-labeled 17β-hydroxy-17αmethyl- estra-4,9, ll-trien-3-one) binds to both androgen and progestin receptors, and consequently, the binding of this ligand to the androgen receptor was assessed in the presence of a 500-fold excess of triamcinolone acetonide. The amounts of 9S binding (7.8 and 5.8 fmol/mg protein) are similar for dihydrotestosterone and methyltrienolone. The amount of 9S binding of testosterone to thereceptor was also similar to that of dihydrotestosterone, but the affinity of testosterone for the 9S receptor was only a fifth or less of that for dihydrotestosterone. The observation that testosterone binds less avidly than dihydrotestosterone to the receptor may explain the role of dihydrotestosterone formation in androgen physiology.

Original languageEnglish (US)
Pages (from-to)173-176
Number of pages4
JournalJournal of Clinical Endocrinology and Metabolism
Volume56
Issue number1
DOIs
StatePublished - 1983

Fingerprint

Dihydrotestosterone
Androgen Receptors
Cytosol
Testosterone
Prostate
Globulins
Metribolone
Androgens
Progesterone Receptors
Promegestone
Trientine
Triamcinolone Acetonide
Steroid hormones
Density Gradient Centrifugation
Tissue
Steroid Receptors
Concanavalin A
Centrifugation
Physiology
Sucrose

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Characterization of the cytosol androgen receptor of the human prostate. / Wilbert, Dirk M.; Griffin III, Jim; Wilson, Jean D.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 56, No. 1, 1983, p. 173-176.

Research output: Contribution to journalArticle

@article{8f0e43049ff14b788506f20c6cab4b76,
title = "Characterization of the cytosol androgen receptor of the human prostate",
abstract = "Direct measurement of the binding of endogenous androgens to the androgen receptor of human tissues has not been possible because of contamination of tissue with traces of plasma proteins, such as testosterone-binding globulin (TeBG), that contain more androgen-binding capacity than does the receptor itself. Molybdate is known to stabilize the 8–9S forms of other steroid hormone receptors. We took advantage of this phenomenon tocharacterize the androgen receptor of hyperplastic prostates removed at surgery, using sucrose density gradient centrifugation in a vertical rotor. In 10 mM sodium molybdate, theandrogen receptor sediments as a distinct 9.2 ± 0.5S moiety, easily separable from TeBG. Unlike TeBG, the 9S receptor is not removed by absorption with Concanavalin A. [3H]Dihydrotestosterone (3H-labeled 17β-hydroxy-5α-androstan- 3-one) bindingto the 9S receptor is not competed for by excess triamcinoloneacetonide (9α-fluoro-llβ,16α,17α,21-tetrahydroxypregna- l,4-diene-3,20-dione cyclic 16,17-acetonide) or promegestone (17,21-dimethyl-19-non-pregna-4,9-diene-3,20- dione), which are known to bind to the progestin receptor. In contrast, [3H]methyltrienolone (3H-labeled 17β-hydroxy-17αmethyl- estra-4,9, ll-trien-3-one) binds to both androgen and progestin receptors, and consequently, the binding of this ligand to the androgen receptor was assessed in the presence of a 500-fold excess of triamcinolone acetonide. The amounts of 9S binding (7.8 and 5.8 fmol/mg protein) are similar for dihydrotestosterone and methyltrienolone. The amount of 9S binding of testosterone to thereceptor was also similar to that of dihydrotestosterone, but the affinity of testosterone for the 9S receptor was only a fifth or less of that for dihydrotestosterone. The observation that testosterone binds less avidly than dihydrotestosterone to the receptor may explain the role of dihydrotestosterone formation in androgen physiology.",
author = "Wilbert, {Dirk M.} and {Griffin III}, Jim and Wilson, {Jean D.}",
year = "1983",
doi = "10.1210/jcem-56-1-113",
language = "English (US)",
volume = "56",
pages = "173--176",
journal = "Journal of Clinical Endocrinology and Metabolism",
issn = "0021-972X",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Characterization of the cytosol androgen receptor of the human prostate

AU - Wilbert, Dirk M.

AU - Griffin III, Jim

AU - Wilson, Jean D.

PY - 1983

Y1 - 1983

N2 - Direct measurement of the binding of endogenous androgens to the androgen receptor of human tissues has not been possible because of contamination of tissue with traces of plasma proteins, such as testosterone-binding globulin (TeBG), that contain more androgen-binding capacity than does the receptor itself. Molybdate is known to stabilize the 8–9S forms of other steroid hormone receptors. We took advantage of this phenomenon tocharacterize the androgen receptor of hyperplastic prostates removed at surgery, using sucrose density gradient centrifugation in a vertical rotor. In 10 mM sodium molybdate, theandrogen receptor sediments as a distinct 9.2 ± 0.5S moiety, easily separable from TeBG. Unlike TeBG, the 9S receptor is not removed by absorption with Concanavalin A. [3H]Dihydrotestosterone (3H-labeled 17β-hydroxy-5α-androstan- 3-one) bindingto the 9S receptor is not competed for by excess triamcinoloneacetonide (9α-fluoro-llβ,16α,17α,21-tetrahydroxypregna- l,4-diene-3,20-dione cyclic 16,17-acetonide) or promegestone (17,21-dimethyl-19-non-pregna-4,9-diene-3,20- dione), which are known to bind to the progestin receptor. In contrast, [3H]methyltrienolone (3H-labeled 17β-hydroxy-17αmethyl- estra-4,9, ll-trien-3-one) binds to both androgen and progestin receptors, and consequently, the binding of this ligand to the androgen receptor was assessed in the presence of a 500-fold excess of triamcinolone acetonide. The amounts of 9S binding (7.8 and 5.8 fmol/mg protein) are similar for dihydrotestosterone and methyltrienolone. The amount of 9S binding of testosterone to thereceptor was also similar to that of dihydrotestosterone, but the affinity of testosterone for the 9S receptor was only a fifth or less of that for dihydrotestosterone. The observation that testosterone binds less avidly than dihydrotestosterone to the receptor may explain the role of dihydrotestosterone formation in androgen physiology.

AB - Direct measurement of the binding of endogenous androgens to the androgen receptor of human tissues has not been possible because of contamination of tissue with traces of plasma proteins, such as testosterone-binding globulin (TeBG), that contain more androgen-binding capacity than does the receptor itself. Molybdate is known to stabilize the 8–9S forms of other steroid hormone receptors. We took advantage of this phenomenon tocharacterize the androgen receptor of hyperplastic prostates removed at surgery, using sucrose density gradient centrifugation in a vertical rotor. In 10 mM sodium molybdate, theandrogen receptor sediments as a distinct 9.2 ± 0.5S moiety, easily separable from TeBG. Unlike TeBG, the 9S receptor is not removed by absorption with Concanavalin A. [3H]Dihydrotestosterone (3H-labeled 17β-hydroxy-5α-androstan- 3-one) bindingto the 9S receptor is not competed for by excess triamcinoloneacetonide (9α-fluoro-llβ,16α,17α,21-tetrahydroxypregna- l,4-diene-3,20-dione cyclic 16,17-acetonide) or promegestone (17,21-dimethyl-19-non-pregna-4,9-diene-3,20- dione), which are known to bind to the progestin receptor. In contrast, [3H]methyltrienolone (3H-labeled 17β-hydroxy-17αmethyl- estra-4,9, ll-trien-3-one) binds to both androgen and progestin receptors, and consequently, the binding of this ligand to the androgen receptor was assessed in the presence of a 500-fold excess of triamcinolone acetonide. The amounts of 9S binding (7.8 and 5.8 fmol/mg protein) are similar for dihydrotestosterone and methyltrienolone. The amount of 9S binding of testosterone to thereceptor was also similar to that of dihydrotestosterone, but the affinity of testosterone for the 9S receptor was only a fifth or less of that for dihydrotestosterone. The observation that testosterone binds less avidly than dihydrotestosterone to the receptor may explain the role of dihydrotestosterone formation in androgen physiology.

UR - http://www.scopus.com/inward/record.url?scp=84995972556&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84995972556&partnerID=8YFLogxK

U2 - 10.1210/jcem-56-1-113

DO - 10.1210/jcem-56-1-113

M3 - Article

C2 - 6183286

AN - SCOPUS:84995972556

VL - 56

SP - 173

EP - 176

JO - Journal of Clinical Endocrinology and Metabolism

JF - Journal of Clinical Endocrinology and Metabolism

SN - 0021-972X

IS - 1

ER -