The use of deoxyribonucleic acid-cellulose chromatography and isoelectric focusing for the characterization and partial purification of steroid-receptor complexes.
The use of deoxyribonucleic acid-cellulose chromatography and isoelectric focusing for the characterization and partial purification of steroid-receptor complexes.
复制标题
使用脱氧核糖核酸-纤维素色谱和等电聚焦来表征和部分纯化类固醇-受体复合物。
DOI：
10.1042/bj1340113
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发表时间：
1973
期刊：
影响因子：
--
通讯作者：
Androgen
中科院分区：
文献类型：
--
作者：
B. W. I. P. Mainwaring;R. Irving;Androgen
1.特异的高亲和力类固醇结合蛋白或受体的两个特性，即它们与DNA-纤维素结合的能力和相对酸性的等电点，已被用作纯化手段。这两个基本特性将受体区别于血清中的类固醇结合蛋白和激素反应细胞中的非特异性低亲和力类固醇结合蛋白。2.通过这些步骤，无论是胞质还是核内的类固醇受体复合体都能获得相当程度的纯化。受体复合体的纯度足以使将来研究它们对代谢过程的可能控制。3.经广泛纯化后，胞质雄激素-受体复合体的沉降系数等物理化学性质没有变化。此外，纯化的复合体完全保留了其至少一个基本的生理特性，即在体外将5α-二氢睾酮(17β-羟基-5α-雄烷-3-酮)转移到染色质中的能力。4.这些方法也可用于研究受体复合体的结构和性质的变化，这是细胞质受体复合体向核染色质转移的必要前提。雄激素受体复合体与染色质结合的温度依赖性本质上是由于细胞质受体复合体在与核染色质结合之前发生了重大变化。5.这些分析程序的分辨率足以对不同男性附腺的受体蛋白进行关键比较。从这些研究中，没有实质性的证据支持雄激素受体的组织特异性；相反，来自不同雄激素依赖腺的受体在物理化学性质上非常相似。6.虽然这些方法最初是用于雄激素受体复合体的部分纯化，但它们同样适用于雌激素受体和孕激素受体复合体的快速和广泛的纯化。
1. Two characteristic properties of the specific high-affinity steroid-binding proteins or receptors, their ability to bind to DNA-cellulose and their relatively acidic isoelectric point, have been exploited as a means of purification. These two fundamental properties distinguish the receptors from the steroid-binding proteins in serum and the non-specific low-affinity steroid-binding proteins in hormone-responsive cells. 2. A significant degree of purification of both cytoplasmic and nuclear steroid-receptor complexes can be achieved with practical facility by these procedures. The purity of the receptor complexes is sufficient to enable studies on their possible control of metabolic processes to be investigated in the future. 3. After extensive purification the physicochemical properties of the cytoplasmic androgen-receptor complex, such as sedimentation coefficient, were unchanged. Further, the purified complex fully retained at least one of its fundamental physiological properties, namely the ability to transfer 5alpha-dihydrotestosterone (17beta-hydroxy-5alpha-androstan-3-one) into chromatin in vitro. 4. The methods may also be employed for studying the changes in the structure and properties of the receptor complexes that are an essential prerequisite for the transfer of cytoplasmic receptor complexes into nuclear chromatin. The temperature-dependence of the binding of androgen-receptor complexes into chromatin is essentially due to a major change in cytoplasmic receptor complex before its attachment to nuclear chromatin. 5. The resolution of these analytical procedures was sufficient to enable a critical comparison of the receptor proteins from different male accessory glands to be undertaken. From these studies, no substantial evidence in support of the tissue specificity of androgen receptors could be established; rather the receptors from different androgen-dependent glands were remarkably similar in physicochemical properties. 6. Although the methods were initially developed for the partial purification of androgen-receptor complexes, they are equally suitable for the prompt and extensive purification of oestrogen-receptor and progesterone-receptor complexes.