Partial purification of nuclear androgen receptor by micrococcal nuclease digestion of chromatin and hydrophobic interaction chromatography.
Partial purification of nuclear androgen receptor by micrococcal nuclease digestion of chromatin and hydrophobic interaction chromatography.
复制标题
通过微球菌核酸酶消化染色质和疏水相互作用层析部分纯化核雄激素受体。
DOI：
10.1111/j.1432-1033.1981.tb05717.x
复制
发表时间：
1981
期刊：
影响因子：
--
通讯作者：
Trudy Comeau
中科院分区：
文献类型：
--
作者：
N. Bruchovsky;P. Rennie;Trudy Comeau
微球菌核酸酶广泛（20%）消化前列腺染色质连接DNA导致95%的核雄激素受体沉淀。将富集受体的沉淀物溶解于pH 7.0，含0.6—1.2 M NaCl的Tes缓冲液中，采用疏水相互作用色谱法进行分析。受体对琼脂糖-氨基-烷基衍生物的吸附随着烷基取代基的长度而增加，表明受体分子表面存在疏水区域。酶切连接体DNA，然后用5-氨基己基琼脂糖对沉淀的染色质蛋白进行层析，受体的平均纯化率为93倍，回收率为45%。这种核雄激素受体部分分离的方法可能与亲和层析等更有选择性的纯化技术相结合。
Extensive (20%) digestion of linker DNA of prostatic chromatin with micrococcal nuclease resulted in the precipitation of 95% of the nuclear androgen receptors. The receptor-enriched precipitate was dissolved in Tes buffer, pH 7.0, containing 0.6--1.2 M NaCl and analysed by hydrophobic interaction chromatography. The adsorption of receptor to omega-amino-alkyl derivatives of agarose increased with the length of the alkyl substituent indicating the presence of hydrophobic regions on the surface of the receptor molecule. Digestion of linker DNA followed by chromatography of precipitated chromatin proteins using 5-aminohexyl-agarose gave rise to a mean 93-fold purificaton of receptor with a recovery of 45%. This approach to the partial separation of nuclear androgen receptor may prove useful in conjunction with more selective purification techniques such as affinity chromatography.