Goueli S A, Holtzman J L, Ahmed K
Biochem Biophys Res Commun. 1984 Sep 17;123(2):778-84. doi: 10.1016/0006-291x(84)90297-3.
The androgen receptor was purified from rat ventral prostate. The purified receptor migrated as a single band of mol. wt. 87000 on SDS-polyacrylamide gels, had a kd for R-1881 (17 beta-hydroxy-17 alpha-methyl-estra-4,9,11-trien-3-one) binding as 6 nM, and sedimentation coefficient of 4.5 S. Phosphorylation of the purified receptor was studied by incubating it with [gamma-32P]ATP in the presence of several purified protein kinases including cAMP-dependent protein kinase, and four cAMP-independent protein kinases (which were active towards substrates such as phosvitin and casein). Phosphorylation of the 87000 mol. wt. androgen receptor protein occurred only in the presence of a nuclear cAMP-independent protein kinase (of the N2 type). No auto-phosphorylation of the receptor was detected. The results indicate that the androgen receptor is a phosphoprotein. Further, phosphorylation of the androgen receptor by only a specific nuclear cAMP-independent protein kinase may be important in determining the dynamics of its function.
雄激素受体是从大鼠腹侧前列腺中纯化出来的。纯化后的受体在十二烷基硫酸钠-聚丙烯酰胺凝胶上迁移为一条分子量为87000的单带，与R-1881（17β-羟基-17α-甲基-雌甾-4,9,11-三烯-3-酮）结合的解离常数为6 nM，沉降系数为4.5 S。通过在几种纯化的蛋白激酶（包括cAMP依赖性蛋白激酶和四种对底物如卵黄高磷蛋白和酪蛋白有活性的cAMP非依赖性蛋白激酶）存在的情况下，将纯化后的受体与[γ-32P]ATP一起孵育，研究其磷酸化情况。分子量为87000的雄激素受体蛋白的磷酸化仅在一种核cAMP非依赖性蛋白激酶（N2型）存在时发生。未检测到受体的自磷酸化。结果表明雄激素受体是一种磷蛋白。此外，仅由一种特定的核cAMP非依赖性蛋白激酶对雄激素受体进行磷酸化，可能在决定其功能动态方面很重要。
