Chang C H, Lobl T J, Rowley D R, Tindall D J
Biochemistry. 1984 May 22;23(11):2527-33. doi: 10.1021/bi00306a032.
An androgen affinity label, 17 beta-[(bromoacetyl)-oxy]-5 alpha-androstan-3-one, has been synthesized in both radioactive and nonradioactive forms. The affinity label (170 Ci/mmol) was characterized and found to have a high degree of purity. Affinity labeling of the androgen receptor from rat ventral prostate was androgen specific and appeared to be directed at the steroid binding site of the protein. Covalent binding was achieved at 0 degrees C; however, heat treatment at 23 degrees C for 30 min enhanced covalent binding by 31%. The covalently bound steroid was resistant to extraction with organic solvents and precipitation with trichloracetate. The Stokes radius (4.2 nm) and sedimentation coefficient (4.5 S) were identical with those found for receptor bound noncovalently to dihydrotestosterone. Gel electrophoresis of the affinity-labeled receptor under denaturing conditions revealed a molecular weight of 86000. The same molecular weight was observed for the receptor from rat seminal vesicle. This affinity label will be useful in future studies on the structure and function of androgen receptors.
已合成了一种雄激素亲和标记物，即17β - [(溴乙酰基)-氧基]-5α - 雄甾烷-3 - 酮，有放射性和非放射性两种形式。该亲和标记物（170居里/毫摩尔）经鉴定具有高度纯度。对大鼠腹侧前列腺雄激素受体进行的亲和标记具有雄激素特异性，且似乎针对该蛋白质的类固醇结合位点。在0℃时实现了共价结合；然而，在23℃下热处理30分钟可使共价结合增加31%。共价结合的类固醇对有机溶剂萃取和三氯乙酸沉淀具有抗性。斯托克斯半径（4.2纳米）和沉降系数（4.5 S）与非共价结合二氢睾酮的受体的相应值相同。在变性条件下对亲和标记的受体进行凝胶电泳，显示分子量为86000。大鼠精囊的受体也观察到相同的分子量。这种亲和标记物将有助于未来对雄激素受体结构和功能的研究。
