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Characterization of androgen receptor by high performance liquid chromatography and sucrose density gradient
M Koutsilieris1, F Grondin, F Radwan
1Department of Biochemistry, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
Abstract:
In the present study we have characterized androgen-receptor complexes of normal and malignant human prostate cytosols using sucrose density gradient centrifugation, gel permeation and anion exchange high performance liquid chromatography (HPLC). Our results indicated that: 1) malignant tissue cytosols differed from normal by the presence of a 4-5S androgen receptor form which accounted for 30% of total specific-binding of malignant tissue cytosols, 2) 8-9S androgen-receptor complexes in normal and malignant prostate cytosols were estimated as 270kDa by gel permeation HPLC, 3) 8-9S complexes were retained and could be eluted by 0.22M KCl on a linear gradient anion exchange HPLC, 4) 4-5S androgen-receptor complexes were estimated as 90kDa by gel permeation HPLC and were not retained on anion exchange HPLC in our experimental conditions, and 5) either 10X dilution of the 4-5S complexes and subsequent anion exchange HPLC, or anion exchange chromatography of 8-9S complexes at 22 degrees C were causing fragmentation of the androgen receptor molecule from normal and malignant tissues. These fragments had enhanced affinity for anion exchange columns. These results are discussed in relation to the composition of the nontransformed androgen receptor macromolecule.