Greene G L, Closs L E, Fleming H, DeSombre E R, Jensen E V
Proc Natl Acad Sci U S A. 1977 Sep;74(9):3681-5. doi: 10.1073/pnas.74.9.3681.
Immunoglobulin obtained from the serum of rabbits immunized with a highly purified preparation of estradiol-receptor complex from calf uterine nuclei has been shown to contain specific antibodies to the receptor protein (estrophilin) by four criteria: (a) precipitation of the radioactive steroid upon addition of goat antibody against rabbit gamma globulin to a mixture of the tritiated estradiol-receptor complex and the immunoglobulin, (b) adsorption of the estradiol-receptor complex by the immunoglobulin linked to Sepharose, (c) adsorption of the estradiol-receptor complex in the presence of the immunoglobulin by Staphylococcus aureus protein-A linked to Sepharose, and (d) the ability of the immunoglobulin to increase the sedimentation rate of the estradiol-receptor complex. Antibodies to calf nuclear estrophilin were shown to crossreact with the nuclear receptor of rat uterus, as well as with the extranuclear receptor of calf, rat, mouse, and guinea pig uterus and of human breast cancer. The antibodies do not react with either the nuclear or extranuclear dihydrotestosterone-receptor complexes of rat prostate or with the extranuclear progesterone-receptor complex of chick oviduct. These findings indicate an immunochemical similarity among estrophilins from several mammalian species, as well as between extranuclear and nuclear forms of the receptor, but not among receptor proteins for different steroid hormones.
从用来自小牛子宫核的高度纯化的雌二醇 - 受体复合物制剂免疫的兔子血清中获得的免疫球蛋白,已通过四个标准显示含有针对受体蛋白(雌激素受体蛋白)的特异性抗体:(a)向氚化雌二醇 - 受体复合物与免疫球蛋白的混合物中加入抗兔γ球蛋白的山羊抗体后,放射性类固醇沉淀;(b)与琼脂糖偶联的免疫球蛋白对雌二醇 - 受体复合物的吸附;(c)与琼脂糖偶联的金黄色葡萄球菌蛋白A在免疫球蛋白存在下对雌二醇 - 受体复合物的吸附;以及(d)免疫球蛋白增加雌二醇 - 受体复合物沉降速率的能力。已证明针对小牛核雌激素受体蛋白的抗体与大鼠子宫的核受体以及小牛、大鼠、小鼠和豚鼠子宫及人乳腺癌的核外受体发生交叉反应。这些抗体不与大鼠前列腺的核内或核外二氢睾酮 - 受体复合物或鸡输卵管的核外孕酮 - 受体复合物发生反应。这些发现表明几种哺乳动物物种的雌激素受体蛋白之间以及受体的核外和核内形式之间存在免疫化学相似性,但不同类固醇激素的受体蛋白之间不存在这种相似性。