Hormone regulation of the rodent Harderian gland: binding properties of the androgen receptor in the male golden hamster

1987 ◽  
Vol 112 (1) ◽  
pp. 3-8 ◽  
Author(s):  
F. Vilchis ◽  
A. Hernandez ◽  
A. E. Perez ◽  
G. Perez-Palacios
Keyword(s):  
Androgen Receptor ◽  
Binding Capacity ◽  
Sedimentation Coefficient ◽  
Harderian Gland ◽  
Hormone Regulation ◽  
Apparent Dissociation Constant ◽  
Receptor Complexes ◽  
Male Golden Hamster ◽  
Sucrose Gradient Ultracentrifugation ◽  
Androgen Binding

ABSTRACT Studies were conducted in castrated golden hamsters to assess whether sexual dimorphism and sensitivity to sex steroid hormones in the rodent Harderian gland are mediated by an interaction of androgens with specific intracellular receptors. Physical properties, binding kinetics and stereospecificity of the androgen receptor were analysed using [3H]mibolerone as the radioligand. The presence of [3H]mibolerone–androgen receptor complexes with a sedimentation coefficient of 7–8S was demonstrated in Harderian gland cytosol by a linear sucrose gradient ultracentrifugation technique using a vertical rotor. Kinetic analysis revealed an androgen-binding site with an apparent dissociation constant of 0·3±0·07 (s.d.) nmol/l and a saturation binding capacity of 113±15 fmol/mg protein. Displacement studies indicated that unlabelled mibolerone, methyltrienolone, 5α-dihydrotestosterone and testosterone were efficient competitors for the androgen-binding sites, while progesterone, 17β-oestradiol, dexamethasone, dehydroepiandrosterone, ethiocholanolone and 5α-16-androsten-3-one were not. Experiments in long-term castrated animals revealed that the Harderian gland androgen receptor concentration and sedimentation coefficient remained unmodified. The results of these studies were interpreted as demonstrating the presence of a specific high-affinity intracellular androgen receptor in the male hamster Harderian gland. J. Endocr. (1987) 112, 3–8

Steroid hormone receptors and the sexual phenotype of the Harderian gland in hamsters

1989 ◽  
Vol 121 (1) ◽  
pp. 149-156 ◽  
Author(s):  
F. Vilchis ◽  
G. Pérez-Palacios
Keyword(s):  
Androgen Receptor ◽  
Binding Site ◽  
Hormone Receptors ◽  
Steroid Hormone ◽  
Sedimentation Coefficient ◽  
Harderian Gland ◽  
Steroid Hormone Receptors ◽  
Progestin Receptors ◽  
Female Gland ◽  
Androgen Binding

ABSTRACT To investigate the participation of intracellular steroid hormone receptors in the sexual transformation process of the Harderian gland, a series of experiments were undertaken in adult golden hamsters. The in-vitro labelling of cytosolic steroid-binding sites with appropriate radioligands revealed the presence of androgen, oestrogen and glucocorticoid but not progestin receptors in the glands from animals of both sexes. The androgen receptor of the female gland was further characterized because it was found to be the predominant intracellular steroid receptor. Studies of binding kinetics using [3H]7α,17α-dimethyl-17β-hydroxy-4-oestren-3-one (DMNT) as ligand, demonstrated a high affinity androgen-binding site with an apparent dissociation constant (Kd) of 0·7 nmol/l and maximal saturation binding capacity of 84·0 ± 3·0 (s.d.) fmol/mg protein. Specificity of the androgen receptor was assessed by displacement analysis; DMNT, 5α-dihydrotestosterone, testosterone and 3α-androstanediol were efficient competitors for the androgen-binding site, while oestradiol-17β, progesterone and dexamethasone exhibited very little, if any, competitive potency. The sedimentation coefficient of the androgen receptor in sucrose density gradients was 8–9 S. These data indicate that the physicochemical characteristics of the androgen receptor from the female gland are similar to those previously described in the male gland. The striking observation of a complete lack of oestrogen-inducible and oestrogen-insensitive progestin receptors in glands cytosol, even after stimulation with cholera toxin, adds further support to the concept that the androgen receptor is the key molecule mediating the hormone-induced sexual transformation of the Harderian gland in this species. Journal of Endocrinology (1989) 121, 149–156

Extraction of androgen–receptor complexes from regions of rat ventral prostate nuclei sensitive or resistant to nucleases

1983 ◽  
Vol 99 (1) ◽  
pp. 51-61 ◽  
Author(s):  
P. Davies
Keyword(s):  
Androgen Receptor ◽  
Binding Sites ◽  
Micrococcal Nuclease ◽  
Ventral Prostate ◽  
Receptor Complex ◽  
Sedimentation Analysis ◽  
Receptor Complexes ◽  
Rat Ventral Prostate ◽  
Profile Characteristic ◽  
Androgen Binding

Rat ventral prostate nuclei contain androgen-binding sites which are susceptible or resistant to excision by endonucleolytic action. Those which were susceptible were associated both with oligonucleosomal and subnucleosomal particles. The sedimentation profile characteristic of a nuclear androgen-receptor complex could be obtained by exhaustive nucleolytic digestion or by treatment of fractions with KCl (0·6 mol/l). Androgen-binding sites resistant to DNAase I were also resistant to KCl, whereas those sites resistant to micrococcal nuclease were partially extractable with KCl. Nuclease-resistant sites could be extracted with heparin (10 mg/ml). Androgen–receptor complexes obtained from nuclease-sensitive or nuclease-resistant regions by extraction with KCl or heparin were indistinguishable by routine sedimentation analysis.

ANDROGEN RECEPTORS IN PROLACTIN PRODUCING PITUITARY TUMOURS IN RATS

1977 ◽  
Vol 86 (2) ◽  
pp. 288-298 ◽  
Author(s):  
Arne Attramadal ◽  
Oddvar Naess ◽  
Egil Haug ◽  
Vidar Hansson ◽  
Ken Purvis
Keyword(s):  
Androgen Receptor ◽  
Binding Sites ◽  
Androgen Receptors ◽  
Sedimentation Coefficient ◽  
Ventral Prostate ◽  
Scatchard Analysis ◽  
Receptor Complex ◽  
Pituitary Tumours ◽  
High Affinity ◽  
Androgen Binding

ABSTRACT The androgen receptor system in prolactin secreting oestrogen induced pituitary tumours has been studied. The tumour cytosol was found to contain specific androgen receptors binding [3H]5α-dihydrotestosterone (DHT) and [3H] testosterone (T) with high affinity and low capacity. Scatchard analysis of the saturation data for T revealed one class of high affinity binding sites. The equilibrium constant of dissociation (Kd) was ∼ 4 × 10−10 m and the number of binding sites was calculated to be 12.8 femtomoles/mg protein. The sedimentation coefficient of the androgen receptor complex in low salt sucrose gradients was ∼ 7 S, the electrophoretic mobility (RF) in 3.25 % polyacrylamide gels ∼ 0.5 and the isoelectric point 5.8. The protein nature of the receptor was indicated by the finding that protease, but not DNase and RNase, eliminated androgen binding. Furthermore, the receptor was thermolabile and functionally dependent on free SH-groups since androgen binding was eliminated by heating 45°C for 30 min) and treatment with p-chloromercuriphenyl sulphonate (1 mm). Steroid specificity was tested in vitro by examining the competing efficiency of different unlabelled steroids for the binding of [3H]T. The affinity of DHT for the receptor was approximately twice that of testosterone while the binding affinity of oestradiol-17β and progesterone was very low. Cortisol had no affinity for the androgen receptor. The dissociation of the androgen receptor complex was very slow at 0°C (t ½ > 48 h). Thus, the characteristics of the cytoplasmic androgen receptors of the prolactin producing pituitary tumours are very similar to those of the androgen receptors earlier demonstrated in the anterior pituitary, hypothalamus, ventral prostate, epididymis and testis. The presence of specific androgen receptors in prolactin producing pituitary tumours indicates that androgen is involved in the regulation of synthesis and release of prolactin.

Androgen receptor in the Harderian gland of Rana esculenta

1991 ◽  
Vol 129 (2) ◽  
pp. 227-232 ◽  
Author(s):  
M. d'Istria ◽  
G. Chieffi-Baccari ◽  
L. Di Matteo ◽  
S. Minucci ◽  
B. Varriale ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Binding Capacity ◽  
Specific Binding ◽  
Secretory Activity ◽  
Harderian Gland ◽  
Nuclear Extract ◽  
Rana Esculenta ◽  
Binding Activity ◽  
Single Class ◽  
Nuclear Extracts

ABSTRACT An androgen receptor has been identified in the cytosolic and nuclear extracts of the Harderian gland of the frog, Rana esculenta. A single class of high-affinity binding sites was found: Kd = 1·9±1·3 (s.d.) nmol/l (n = 26) for the cytosolic extract and Kd = 0·9±0·8 nmol/l (n = 15) for the nuclear extract. The presence of binding activity in both nuclear and cytosolic extracts and the low rate of ligand-receptor dissociation are characteristics that distinguish this receptor from a steroid-binding protein. The Kd did not show any sex difference and did not exhibit any secretory activity-related change. Binding in both cytosolic and nuclear extracts was specific for androgens (testosterone = 5α-dihydrotestosterone); oestradiol-17β showed a 30% cross-reaction; moreover, specific binding of [3H]oestradiol-17β was not detectable. The binding capacity of the Harderian gland increased progressively in both fractions from October to December, reaching a peak in May, and decreased suddenly during July to August. The lack of any morphological sex-related difference in the Harderian gland of the green frog might be accounted for by the high amount of circulating androgens as well as a similar concentration of androgen receptor in both sexes. Journal of Endocrinology (1991) 129, 227–232

A SOLUBLE ANDROGEN RECEPTOR IN THE CYTOPLASM OF RAT PROSTATE

1969 ◽  
Vol 45 (4) ◽  
pp. 531-541 ◽  
Author(s):  
W. I. P. MAINWARING
Keyword(s):  
Molecular Weight ◽  
Androgen Receptor ◽  
Nuclear Receptor ◽  
Sedimentation Coefficient ◽  
Binding Specificity ◽  
Supernatant Fraction ◽  
Soluble Receptor ◽  
Tryptophan Residues ◽  
Rat Prostate ◽  
Androgen Binding

SUMMARY A thermolabile protein with the properties of a steroid 'receptor' was identified in the cytoplasmic or 105,000 g supernatant fraction of the rat prostate. The receptor has a particular binding specificity towards 5αdihydrotestosterone. Testosterone is bound to a lesser extent but other steroids, including certain androgenic hormones, are not bound. The sedimentation coefficient of 8·0 s and the frictional ratio of 1·96, equivalent to a molecular weight of 2·74 × 105, clearly distinguish the soluble androgen-receptor from the androgen-binding globulin in serum and the androgen-receptor in the prostatic nucleus. Like the nuclear receptor, however, the soluble receptor is probably an acidic protein. Both cysteine and tryptophan residues appear necessary for maintaining the functional configuration of the receptor.

scholarly journals Development of an Androgen Receptor Inhibitor Targeting the N-Terminal Domain of Androgen Receptor for Treatment of Castration Resistant Prostate Cancer

Cancers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 3488
Author(s):  
Fuqiang Ban ◽  
Eric Leblanc ◽  
Ayse Derya Cavga ◽  
Chia-Chi Flora Huang ◽  
Mark R. Flory ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Androgen Receptor ◽  
Splice Variants ◽  
Full Length ◽  
Cancer Resistance ◽  
Castration Resistant Prostate Cancer ◽  
Terminal Domain ◽  
Castration Resistant ◽  
Coregulatory Proteins ◽  
Androgen Binding

Prostate cancer patients undergoing androgen deprivation therapy almost invariably develop castration-resistant prostate cancer. Resistance can occur when mutations in the androgen receptor (AR) render anti-androgen drugs ineffective or through the expression of constitutively active splice variants lacking the androgen binding domain entirely (e.g., ARV7). In this study, we are reporting the discovery of a novel AR-NTD covalent inhibitor 1-chloro-3-[(5-([(2S)-3-chloro-2-hydroxypropyl]amino)naphthalen-1-yl)amino]propan-2-ol (VPC-220010) targeting the AR-N-terminal Domain (AR-NTD). VPC-220010 inhibits AR-mediated transcription of full length and truncated variant ARV7, downregulates AR response genes, and selectively reduces the growth of both full-length AR- and truncated AR-dependent prostate cancer cell lines. We show that VPC-220010 disrupts interactions between AR and known coactivators and coregulatory proteins, such as CHD4, FOXA1, ZMIZ1, and several SWI/SNF complex proteins. Taken together, our data suggest that VPC-220010 is a promising small molecule that can be further optimized into effective AR-NTD inhibitor for the treatment of CRPC.

Sex difference in the cytosolic and nuclear distribution of androgen receptor in mouse submandibular gland

1986 ◽  
Vol 108 (2) ◽  
pp. 267-273 ◽  
Author(s):  
S. Kyakumoto ◽  
R. Kurokawa ◽  
Y. Ohara-Nemoto ◽  
M. Ota
Keyword(s):  
Androgen Receptor ◽  
Nuclear Receptors ◽  
Binding Sites ◽  
Androgen Receptors ◽  
Dissociation Constants ◽  
Scatchard Analysis ◽  
Male And Female ◽  
Short Period ◽  
Almost All ◽  
Androgen Binding

ABSTRACT Cytosol and nuclear androgen receptors in submandibular glands of male and female mice were measured by an exchange assay at 0 °C. The binding of [3H]methyltrienolone to cytosol receptors in females was mostly saturated within a short period of incubation (3 h), whereas the saturation was much slower in males; suggesting that almost all of the cytosol receptors were unoccupied in females and the receptors were partially occupied in males. Nuclear receptors were extracted with pyridoxal 5′-phosphate (5 mmol/l) from nuclear fractions with 93–95% efficiency. The exchange of the bound steroids occurred by 24–48 h at 0 °C, suggesting that most of the nuclear androgen receptor was occupied. The binding was low at higher temperatures, probably due to inactivation of the receptor. Scatchard analysis showed that the apparent dissociation constants of cytosol and nuclear receptors were similar (0·8 and 0·9 nmol/l respectively) in both sexes. On the other hand, the number of androgen-binding sites in the nucleus was much higher in males than in females (1052 fmol/mg DNA and 32 fmol/mg DNA respectively), while the number in the cytosol was higher in females than in males (512 fmol/mg DNA and 368 fmol/mg DNA respectively). These observations show that androgen receptors exist mainly (74%) in the nuclei of males, while they exist mostly (94%) in the cytosol of females. J. Endocr. (1986) 108, 267–273

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