Koutsilieris M, Bouthillier F, Grondin F, Radwan F, Carmel M, Elhilali M, Lehoux J G
Department of Biochemistry, Faculty of Medicine, University of Sherbrooke, Quebec, Canada.
Prostate. 1988;13(2):155-64. doi: 10.1002/pros.2990130207.
We investigated the application of octadecylsilyl (ODS)-silica in studies related to characterization and purification of the nontransformed rat ventral prostate androgen receptor. The results indicated that ODS-silica successfully separates the free [3H]R1881 from the labeled [3H]R1881 transformed (4-5S) and nontransformed (8-9S) rat ventral prostate androgen receptors. Partial purification of the 8-9S receptor form was performed by the fractionation of rat cytosol using cartridges of the ODS-silica and fast-flow-rate phosphocellulose chromatography. Further purification was accomplished by differential chromatography (DEAE-cellulose and slow-flow-rate phosphocellulose chromatography). This partially purified 8-9S receptor, when analyzed on a gel permeation high-performance liquid chromatography column, resulted in a three-peak pattern of UV absorbance. One of these peaks corresponded to a 59-kD non[3H]R1881-binding protein and the remaining two corresponded to 270-kD and 190-kD[3H]R1881-binding proteins. These results demonstrate the usefulness of ODS-silica in androgen receptor studies. The association of a 59-kD nonsteroid binding protein with the nontransformed rat ventral prostate androgen receptor is discussed.
我们研究了十八烷基硅烷（ODS）-硅胶在非转化大鼠腹侧前列腺雄激素受体的表征和纯化相关研究中的应用。结果表明，ODS-硅胶成功地将游离的[³H]R1881与标记的[³H]R1881转化型（4-5S）和非转化型（8-9S）大鼠腹侧前列腺雄激素受体分离。使用ODS-硅胶柱和快速流速磷酸纤维素色谱法对大鼠胞质溶胶进行分级分离，对8-9S受体形式进行了部分纯化。通过差异色谱法（DEAE-纤维素和慢速流速磷酸纤维素色谱法）完成进一步纯化。当在凝胶渗透高效液相色谱柱上分析这种部分纯化的8-9S受体时，得到了一个三峰的紫外吸收图谱。其中一个峰对应于一种59-kD的非[³H]R1881结合蛋白，其余两个峰对应于270-kD和190-kD的[³H]R1881结合蛋白。这些结果证明了ODS-硅胶在雄激素受体研究中的实用性。讨论了一种59-kD非类固醇结合蛋白与非转化大鼠腹侧前列腺雄激素受体的关联。
