van Laar J H, Bolt-de Vries J, Zegers N D, Trapman J, Brinkmann A O
Department of Biochemistry II, Erasmus University Rotterdam, The Netherlands.
Biochem Biophys Res Commun. 1990 Jan 15;166(1):193-200. doi: 10.1016/0006-291x(90)91930-q.
Androgen receptor heterogeneity and phosphorylation were studied in the human LNCaP cell line. Fluorography after photoaffinity labeling as well as immunoblotting with a specific polyclonal antibody revealed that the human androgen receptor migrated as a closely spaced 110 kD doublet on SDS-polyacrylamide gels. A time-dependent change in the ratio between the two isoforms was not observed after R1881 treatment of intact cells. In nuclear extracts of LNCaP cells that were incubated with [32P]orthophosphate in the presence of 10 nM R1881, a 110 kD phosphorylated protein was demonstrated after immunopurification using a monoclonal antibody against the human androgen receptor. Only a very small amount of this phosphoprotein was detected in the nuclear fraction from cells not treated with R1881. These results indicate that the human androgen receptor in LNCaP cells can be phosphorylated.
在人LNCaP细胞系中研究了雄激素受体的异质性和磷酸化。光亲和标记后的荧光自显影以及用特异性多克隆抗体进行的免疫印迹显示，人雄激素受体在SDS-聚丙烯酰胺凝胶上以紧密间隔的110 kD双峰形式迁移。在用R1881处理完整细胞后，未观察到两种异构体之间比例的时间依赖性变化。在存在10 nM R1881的情况下用[32P]正磷酸盐孵育的LNCaP细胞核提取物中，使用抗人雄激素受体的单克隆抗体进行免疫纯化后，证实了一种110 kD的磷酸化蛋白。在未用R1881处理的细胞的核部分中仅检测到极少量的这种磷蛋白。这些结果表明LNCaP细胞中的人雄激素受体可以被磷酸化。
