Human breast tumor estrogen receptor: effects of molybdate and electrophoretic analyses.
Human breast tumor estrogen receptor: effects of molybdate and electrophoretic analyses.
复制标题
人乳腺肿瘤雌激素受体：钼酸盐和电泳分析的影响。
DOI：
10.1210/endo-108-4-1369
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发表时间：
1981
期刊：
影响因子：
4.8
通讯作者：
M. R. Sherman
中科院分区：
文献类型：
--
作者：
L. Miller;F. Tuazon;E. Niu;M. R. Sherman
最大和最小的离散形式的雌激素受体在人类乳腺肿瘤细胞质的特点是竞争性类固醇结合，ultracentrugation，凝胶过滤，和聚丙烯酰胺凝胶电泳几个浓度。用[~ 3 H]雌二醇孵育胞质溶胶，并在含有20 mM Na_2MoO_4和0或150 mM KCl的甘油梯度中离心，结果显示9- 10 S形式的受体。它类似于其他人类和啮齿动物、恶性和健康组织的细胞质中的酒石酸盐稳定复合物，以及在含有亮抑酶肽（一种细菌蛋白酶抑制剂）的乳腺肿瘤细胞质中检测到的复合物。通过在所有缓冲液中加入β-内酰胺酶，促进了纯化过程中受体完整性的保持和与血清类固醇结合组分的区分。可能的作用机制包括抑制核糖核酸酶对RNA相关受体形式的作用，以及通过稳定脆弱残基或相邻残基上的磷酸基团来保护免受特异性蛋白水解切割。在肿瘤细胞质溶胶的分级分离过程中，在没有受体的情况下，受体被转化为片段的混合物。保留结合类固醇的最小的部分受体类似于内源性和外源性蛋白酶作用于广泛组织中所有类别类固醇的受体的产物。乳腺肿瘤雌激素受体的最大和最小已知形式与其他受体的相应形式之间的相似性支持了类固醇受体在不同来源的正常和恶性组织中的共同结构的概念。
The largest and smallest discrete forms of the estrogen receptor in human breast tumor cytosol were characterized by competitive steroid binding, ultracentrifugation, gel filtration, and electrophoresis in polyacrylamide gels of several concentrations. Incubation of cytosol with [3H]estradiol and centrifugation in glycerol gradients containing 20 mM Na2MoO4 and 0 or 150 mM KCl revealed a 9-10S form of the receptor. It resembles the molybdate-stabilized complexes in cytosols of other human and rodent, malignant and healthy tissues, and the complex detected in breast tumor cytosol containing leupeptin, a bacterial protease inhibitor. Preservation of receptor integrity during purification and discrimination from serum steroid-binding components are facilitated by inclusion of molybdate in all buffers. Possible mechanisms of action of molybdate include the inhibition of ribonuclease action on RNA-associated receptor forms and protection against specific proteolytic cleavage by stabilization of a phosphate group on the vulnerable residue or a neighboring one. During fractionation of tumor cytosol in the absence of molybdate, the receptor is converted to a mixture of fragments. The smallest that retains the bound steroid, the mero-receptor, resembles the products of endogenous and exogenous protease action on receptors for all classes of steroids in a wide range of tissues. The similarities between both the largest and the smallest known forms of the breast tumor estrogen receptor and corresponding forms of other receptors support the notion of the common architecture of steroid receptors in normal and malignant tissues of diverse origins.
DOI：
10.1016/0022-4731(80)90260-5
发表时间：
1980
期刊：
Journal of steroid biochemistry
影响因子：
--
作者：
Grody,WW;Compton,JG;Schrader,WT;O'Malley,BW
通讯作者：
O'Malley,BW