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Sucrose density gradient and exchange high performance liquid chromatography characteristics of rat prostate androgen
M Koutsilieris1, F Grondin, M Carmel
1Department of Biochemistry, University of Sherbrooke, Montreal, Quebec, Canada.
Abstract:
In this study, we characterized rat prostate androgen receptor using sucrose density gradient ultracentrifugation, gel permeation and anion exchange high performance liquid chromatography (HPLC). Our results indicated that 1) androgen receptor complexes in rat prostate cytosols containing no KCl were of 8-9S form corresponding to 270kDa, and in cytosols containing 0.6M KCl were of 5-6S form corresponding to 90kDa; 2) 8-9S androgen receptor complexes were retained and could be eluted by 0.22M KCl in a linear gradient anion exchange HPLC; 3) Dilution (10X) of the 8-9S and of the 5-6S receptor complexes and subsequent chromatography on anion exchange columns was indicative of fragmentation. Fragments were of 3-4S form and were eluted by 0.33M KCl on anion exchange HPLC. Temperature (22 degrees C) was permissive of proteolysis generating a 30kDa androgen receptor fragment. These results are discussed in the context of the composition of non-transformed rat prostate androgen receptor.