Characterization of androgen receptors after photoaffinity labelling with [3H]methyltrienolone (R1881).

Abstract

The synthetic androgen 17 beta-hydroxy-17 alpha-[3H]methyl-4,9,11-estratrien-3-one (R1881) has been used as photoaffinity label to characterize androgen receptors in rat prostate, in a human transplantable prostatic adenocarcinoma (PC-82) and in calf uterus. Androgen receptors preparations were partially purified either via differential chromatography on 2',5'-ADP-Sepharose (rat prostate), via anion exchange fast protein liquid chromatography (rat prostate and PC-82) or via DNA-cellulose chromatography (calf uterus). Purification factors obtained with the three different methods were: 245, 75 and 40 respectively. Photolabelling of receptor preparations was performed via irradiation with a high pressure mercury lamp either before or after partial purification. Polyacrylamide gel electrophoresis under denaturing conditions showed that the DNA-binding form of the androgen receptor in calf uterus cytosol is a protein with a molecular mass of approx 95 kD. The covalent attachment of [3H]R1881 to the 95 kD protein could be completely suppressed by a 200-fold molar excess of dihydrotestosterone. In rat prostate cytosol an androgen receptor with a molecular mass of approx 50 kD could be photoaffinity labelled with R1881. A similar size was found for the androgen receptor in the human prostatic adenocarcinoma. Our results show that photoaffinity labelling of androgen receptors with [3H]R1881 as ligand can be applied for characterization of partial purified androgen receptor preparations.

• Authors: A O Brinkmann, G G Kuiper, W de Boer, E Mulder, J Bolt, G J van Steenbrugge, H J van der Molen
• Journal: Journal of steroid biochemistry (J Steroid Biochem) Vol. 24 Issue 1 Pg. 245-9 (Jan 1986) ISSN: 0022-4731 [Print] England
• PMID: 2422446 (Publication Type: Journal Article)

Chemical References

• Affinity Labels
• Estrenes
• Receptors, Androgen
• Tritium
• Metribolone
• RNA
• DNA

Topics

• Affinity Labels
• Animals
• Cattle
• Cytosol (analysis)
• DNA (metabolism)
• Estrenes
• Female
• Male
• Metribolone
• Photolysis
• Prostate (analysis)
• Prostatic Neoplasms (analysis)
• RNA (metabolism)
• Rats
• Receptors, Androgen (analysis)
• Tritium
• Uterus (analysis)