Characterization of androgen receptors after photoaffinity labelling with [3H]methyltrienolone (R1881).
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.
Brinkmann
A O
AO
Kuiper
G G
GG
de Boer
W
W
Mulder
E
E
Bolt
J
J
van Steenbrugge
G J
GJ
van der Molen
H J
HJ
D000345:Affinity Labels; D000818:Animals; D002417:Cattle; D003600:Cytosol / Q000032:analysis; D004247:DNA / Q000378:metabolism; D004963:Estrenes; D005260:Female; D008297:Male; D015741:Metribolone; D010782:Photolysis; D011467:Prostate / Q000032:analysis; D011471:Prostatic Neoplasms / Q000032:analysis; D012313:RNA / Q000378:metabolism; D051381:Rats; D011944:Receptors, Androgen / Q000032:analysis*; D014316:Tritium; D014599:Uterus / Q000032:analysis
D000345:Affinity Labels; D004963:Estrenes; D011944:Receptors, Androgen; D014316:Tritium; D015741:Metribolone; D012313:RNA; D004247:DNA