Immunoprecipitation of the lutropin receptor. Loss of receptor molecules during down-regulation.
Immunoprecipitation of the lutropin receptor. Loss of receptor molecules during down-regulation.
复制标题
促黄体素受体的免疫沉淀。
DOI：
10.1042/bj2240467
复制
发表时间：
1984
期刊：
影响因子：
--
通讯作者：
H. Rajaniemi
中科院分区：
文献类型：
--
作者：
M. Metsikkö;H. Rajaniemi
用从假孕大鼠卵巢中纯化的促黄体激素受体免疫家兔，制备特异性抗促黄体激素受体抗体。1：100稀释的抗受体血清与抗（兔γ-球蛋白）血清一起免疫沉淀70%的3 H标记的纯化促黄体激素受体和42%的125 I-绒毛膜促性腺激素受体复合物。抗血清抑制激素结合大鼠卵巢颗粒。用高碘酸盐/NaB 3 H4标记假孕大鼠卵巢颗粒，并用Triton X-100溶解。使用抗受体血清对Triton X-100提取物进行免疫沉淀。当免疫沉淀物溶解和分析，在十二烷基硫酸钠聚丙烯酰胺凝胶电泳在还原条件下，然后通过荧光，受体多肽与一个明显的Mr 95000检测。将受体下调剂量的绒毛膜促性腺激素注射到假孕大鼠中，4天后取出它们的卵巢并匀浆，如上分析免疫可沉淀受体。相应地，促黄体激素受体分子实际上消失了，而不仅仅是在同源下调过程中被激素掩盖。
Specific anti-(lutropin receptor) antibodies were produced by immunizing rabbits with lutropin receptor purified from pseudopregnant rat ovary. The anti-receptor serum at 1:100 dilution together with anti-(rabbit gamma-globulin) serum immunoprecipitated 70% of 3H-labelled, purified lutropin receptor and 42% of 125I-chorio-gonadotropin-receptor complex. The antiserum inhibited hormone binding to rat ovarian particles. Pseudopregnant rat ovarian particles were labelled with periodate/NaB3H4 and solubilized with Triton X-100. The Triton X-100 extract was subjected to immunoprecipitation using the anti-receptor serum. When the immunoprecipitate was dissolved and analysed by polyacrylamide-gel electrophoresis in sodium dodecyl sulphate under reducing conditions followed by fluorography, a receptor polypeptide with an apparent Mr 95000 was detected. A receptor down-regulating dose of choriogonadotropin was injected into pseudopregnant rats and their ovaries were removed and homogenized 4 days later, and analysed for immunoprecipitable receptors as above. No receptor molecules were found. Accordingly, the lutropin receptor molecules actually disappear rather than merely become masked from hormone during homologous down-regulation.