Photoaffinity labelling with [3H]R1881 of androgen receptors from human cultured genital fibroblasts of normal individuals and patients with androgen receptor abnormalities

Gyorki, S.; Warne, Garry L.; Funder, John W.

doi:10.1016/0022-4731(88)90159-8

Cited by 6 publications

(5 citation statements)

References 18 publications

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“…We determined that androgen binding was diminished, but not absent, suggesting a qualitative defect of binding. We have also previously shown that partially purified AR from subject Te24 has a reduced molecular weight (M r ∼43 kDa), suggesting this receptor is truncated (Gyorki et al, 1988). The presence of detectable androgen binding in fibroblasts from this subject suggests that the AR is not truncated in vivo.…”

Section: Te24mentioning

confidence: 77%

“…Her testes were removed at age 6 yrs, and a genital skin biopsy was taken at age 12 yrs. (Gyorki et al, 1988). 3 n/d -binding not determined.…”

Section: Subjectsmentioning

confidence: 99%

“…Previous studies performed in our laboratory on fibroblasts from subject Te24 had shown the presence of a truncated receptor protein following photoaffinity labeling and affinity purification (Gyorki et al, 1988). This study used cytosol extracted from genital skin fibroblasts to partially purify the AR via affinity chromatography.…”

Section: Androgen Binding Analysismentioning

confidence: 99%

“…To determine the ligand binding ability of the AR, androgen binding was measured in cultured genital skin fibroblasts (Table 1), as previously described (MacLean et al, 1993). Previous studies performed in our laboratory on fibroblasts from subject Te24 had shown the presence of a truncated receptor protein following photoaffinity labeling and affinity purification (Gyorki et al, 1988). This study used cytosol extracted from genital skin fibroblasts to partially purify the AR via affinity chromatography.…”

Section: Androgen Binding Analysismentioning

confidence: 99%

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Novel androgen receptor gene mutations in Australian patients with complete androgen insensitivity syndrome

et al. 2004

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We have identified androgen receptor (AR) gene mutations in eight Australian subjects with complete androgen insensitivity syndrome (AIS). Four individuals, from three families, have novel mutations that introduce premature termination codons. Two siblings have the nonsense mutation Glu681X, and another subject has the nonsense mutation p.Ser884X. The other subject has a CA insertion at codon 829 (c.2847_2848insCA), causing a frameshift mutation that introduces four nonsense amino acids prior to a Stop codon. All the termination codons occur in the ligand binding domain, and cause reduced androgen binding in patient genital skin fibroblasts. Four further patients have missense mutations. One subject has two different mutations, p.Ala645Asp in the hinge region of the receptor, and p.Arg752Gln in the ligand binding domain. Both these mutations have previously been reported in patients with AIS, but the combination of these two mutations in one subject is unique. Another subject has a novel c.2533G>C transversion at the first nucleotide in exon 5, introducing the amino acid change p.Gly724Ala at a highly conserved residue in the ligand binding domain. Androgen binding is normal in fibroblasts from this subject, although other point mutations at this amino acid totally abolish binding. Two other subjects have mutations previously described as causing AIS, namely p.Arg779Trp and p.Val889Met substitutions in the ligand binding domain of the receptor. The p.Arg779Trp mutation is associated with the detection of a truncated AR protein in this patient's fibroblasts, suggesting the mutation renders the receptor susceptible to proteolysis.

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Section: Te24mentioning

confidence: 77%

“…Her testes were removed at age 6 yrs, and a genital skin biopsy was taken at age 12 yrs. (Gyorki et al, 1988). 3 n/d -binding not determined.…”

Section: Subjectsmentioning

confidence: 99%

Section: Androgen Binding Analysismentioning

confidence: 99%

Section: Androgen Binding Analysismentioning

confidence: 99%

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Novel androgen receptor gene mutations in Australian patients with complete androgen insensitivity syndrome

et al. 2004

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“…2a and 2b). Further information concerning the structure of the AR has become available from photoaffinity labelling with R1881 [34][35][36] and cloning of the gene [5][6][7][8]: the AR protein has a molecular mass ranging from 90 to 110 kDa, with an androgen-binding domain corresponding to a 30 kDa portion of the molecule [36].…”

Section: Discussionmentioning

confidence: 99%

Radiation-inactivation size of transformed and non-transformed androgen receptors

Turcotte

Beauregard

Potier

et al. 1991

Biochemical Journal

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The nucleic acid sequence of the androgen receptor (AR) gene predicts that the protein structure possesses DNA- and steroid-binding domains that show high degrees of sequence similarity with those of other steroid receptors. Since the steroid-binding domain of the AR corresponds to a 30 kDa portion of the protein, and the AR structure may be monomeric or hetero-oligomeric depending on its transformation state, we have herein determined the AR radiation-inactivation size (RIS) in relation to the molecular structure whose binding activity toward methyltrienolone (R1881) is abolished by a radiation 'hit'. Soluble fractions from whole canine prostatic tissue were used as a source of non-transformed AR. The AR transformation was induced by the addition of 0.6 M-KCl, and these preparations were used together with high-salt nuclear extracts as a source of transformed AR. To maximize the binding activity, molybdate and dithiothreitol were included during AR extraction. Receptor transformation was verified by modifications of both the sedimentation coefficients (from 7.5 S to 4.1 S on sucrose gradients) and molecular masses (from 260 kDa to 115 kDa by gel filtration). The RIS values of the non-transformed and transformed ARs were not statistically different: 92 +/- 19 kDa and 110 +/- 25 kDa respectively. In addition, the inactivation of AR binding activity by radiation was attributed to a loss of binding sites, with no significant change in the Kd. When benzoic acid, a free-electron scavenger, was added together with dithiothreitol before and after irradiation, no change in the RIS value was observed. Thus, in the canine prostate, the RIS value of the AR represents the monomeric protein, independently of its association with other proteins, and this value corresponds to that predicted by cloning studies and photoaffinity-labelling of AR.

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Establishment and characterization of monoclonal antibody against androgen receptor

Demura

Kuzumaki

Oda

et al. 1989

Journal of Steroid Biochemistry

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Photoaffinity labelling with [3H]R1881 of androgen receptors from human cultured genital fibroblasts of normal individuals and patients with androgen receptor abnormalities

Cited by 6 publications

References 18 publications

Novel androgen receptor gene mutations in Australian patients with complete androgen insensitivity syndrome

Novel androgen receptor gene mutations in Australian patients with complete androgen insensitivity syndrome

Radiation-inactivation size of transformed and non-transformed androgen receptors

Establishment and characterization of monoclonal antibody against androgen receptor

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