A MOLECULAR APPROACH TO GONADOTROPIN RECEPTOR REGULATION

我们已经成功地开发出溶解黄体生成素/人绒毛膜促性腺激素受体的程序 (LHR)来自猪和大鼠的卵巢，产量高。猪和大鼠LHR有 经两步hCG-琼脂糖亲和层析纯化至表观均一 层析程序。如此纯化的LHR的主要亚基有 变性还原条件下的分子量为7-8万道尔顿 条件。在拟议的资助期内，物理化学和 比较纯化的大鼠和猪LHR的生物学特性。 针对猪LHR的抗体将在兔体内开发，而大鼠LHR将在 注射到小鼠体内，目的是产生一种针对大鼠的单抗 哈哈。抗体将通过免疫化学技术和 它们的生物学特性将使用分离的受体进行测试， 膜、培养细胞和整个动物。的分子基础 激素对受体的调节将通过直接分析 比色法测定LHR的合成和降解速率 免疫沉淀方法学。 受体抗体将为研究提供一个极好的分析工具 LHR合成、加工和周转的分子生物学研究。LHR 抗体不仅将成为研究卵巢疾病的关键工具 在分子水平上的生理学，也可能具有重大的实用价值 在免疫避孕和认识不孕症中的应用 卵巢起源及LHR单抗药物的研制 可能在卵巢癌治疗中有用的结合物。

We have successfully developed procedures to solubilize LH/hCG receptors (LHR) from porcine and rat ovaries in high yield. Porcine and rat LHR have been purified to apparent homogeneity by a two-step hCG-Sepharose affinity chromatographic procedure. The predominant subunit of LHR so purified has a molecular weight of 70,000 - 80,000 Daltons under denaturing reducing conditions. During the proposed grant period, the physico-chemical and biological properties of purified rat and porcine LHR will be compared. Antibodies to porcine LHR will be developed in rabbits, and rat LHR will be injected into mice with the goal of producing a monoclonal antibody to rat LHR. Antibodies will be characterized by immunochemical techniques and their biological properties will be tested using isolated receptors, membranes, cultured cells and the whole animal. The molecular basis of receptor regulation by hormones will be analyzed through direct measurements of rates of synthesis and degradation of LHR by specific immunoprecipitation methodology. Receptor antibodies will provide a superb analytical tool for investigation of the molecular biology of LHR synthesis, processing and turnover. LHR antibodies will not only be critical tools for investigation of ovarian physiology at the molecular level, but also may have significant practical application in immunocontraception and in understanding infertility of ovarian origin and in the development of monoclonal LHR antibody-drug conjugates which may be useful in therapy of ovarian cancer.