Androgen effects on the solubility and conformational change of the androgen receptor in baculovirus expression system.
Androgen effects on the solubility and conformational change of the androgen receptor in baculovirus expression system.
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雄激素影响杆状病毒表达系统中雄激素受体的溶解度和构象变化。
DOI：
10.1023/a:1006906132516
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发表时间：
1999
影响因子：
4.3
通讯作者：
Chang,C
中科院分区：
文献类型：
--
作者：
Wang,C;Young,WJ;Chang,C
为了从杆状病毒表达系统中高效纯化雄激素受体（AR），我们将6个组氨酸残基与AR的N端结构域融合作为标签，与Ni+2亲和柱特异性结合。我们的数据表明，加入雄激素可以增加AR与Ni+2-亲和柱的结合能力，这种AR结合能力的增加可能是由于雄激素诱导的N端结构域的组氨酸残基的暴露。雄激素增强的Ni+2-柱结合也与AR溶解度的增加相关。电泳迁移率变动分析进一步表明，只有纯化的AR才能与雄激素反应元件相互作用。总之，我们的数据表明，雄激素的AR的激素结合结构域的结合可能会导致AR的N-末端结构域的构象变化，并增加AR的亲水性。
To purify the androgen receptor (AR) efficiently from baculovirus expression system, we fused 6 histidine residues with the N-terminal domain of AR as a tag to specifically bind to Ni+2-affinity column. Our data indicated that adding androgen can increase the binding capacity of his-tag AR to the Ni+2-affinity column, and this increased binding capacity of AR could be due to the exposure of histidine residues of N-terminal domain induced by androgen. The androgen-enhanced binding to Ni+2-column also correlated with the increasing solubility of AR. Electrophoretic mobility shift assay further indicated that only purified AR could interact with androgen response element. Together, our data suggest that the binding of androgen to the hormone binding domain of AR may result in the conformational change of the N-terminal domain of AR and increase the hydrophilic property of AR.