MOLECULAR APPROACH TO GONADOTROPIN RECEPTOR REGULATION

促性腺激素受体调节的分子方法

• 批准号: 3320173
• 负责人: JAY wimalasena WIMALASENA
• 金额: $ 10.04万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 1989-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3320173
• 关键词: active immunization; antibody formation; autoradiography; chorionic gonadotropin; chromatography; density gradient ultracentrifugation; electrophoresis; histochemistry /cytochemistry; hormone biosynthesis; hormone receptor; hormone regulation /control mechanism; immunochemistry; laboratory mouse; laboratory rabbit; laboratory rat; luteinizing hormone; monoclonal antibody; ovary; passive immunization; radioassay; swine

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.

我们已经成功地开发了溶解LH/hCG受体的方法 (LHR)从猪和大鼠卵巢中以高产量获得。 猪和大鼠LHR具有 通过两步hCG-琼脂糖亲和层析纯化至表观均一 色谱程序。 如此纯化的LHR的主要亚基具有 在变性还原条件下，分子量为70,000 - 80,000道尔顿 条件 在拟议的赠款期间， 将比较纯化的大鼠和猪LHR的生物学特性。 猪LHR的抗体将在家兔中开发，大鼠LHR将在 注射到小鼠体内，目的是产生抗大鼠的单克隆抗体， LHR 抗体将通过免疫化学技术表征， 将使用分离的受体测试它们的生物学特性， 膜、培养细胞和整个动物。 的分子基础 受体调节激素将分析通过直接 LHR的合成和降解速率的测量 免疫沉淀方法学。 受体抗体将为研究提供极好的分析工具 LHR合成、加工和周转的分子生物学。 LHR 抗体不仅是研究卵巢癌的重要工具， 在分子水平上的生理学，但也可能有重大的实际意义， 在免疫避孕和了解不孕症方面的应用 卵巢起源和单克隆LHR抗体药物的开发 可用于治疗卵巢癌的缀合物。

项目成果

Ethanol has direct inhibitory effects on steroidogenesis in human granulosa cells: specific inhibition of LH action.

• DOI: 10.1111/j.1530-0277.1990.tb01192.x
• 发表时间: 1990-08
• 期刊: Alcoholism, clinical and experimental research
• 作者: S. Saxena;D. Meehan;P. Coney;J. Wimalasena

Biological actions of monoclonal luteinizing hormone/human chorionic gonadotropin receptor antibodies.

单克隆黄体生成激素/人绒毛膜促性腺激素受体抗体的生物学作用。

• DOI: 10.1095/biolreprod46.2.265
• 发表时间: 1992
• 期刊: Biology of reproduction
• 影响因子: 3.6
• 作者: Indrapichate,K;Meehan,D;Lane,TA;Chu,SY;Rao,CV;Johnson,D;Chen,TT;Wimalasena,J
• 通讯作者: Wimalasena,J

Purification and characterization of the human ovarian LH/hCG receptor and comparison of the properties of mammalian LH/hCG receptors.

人卵巢 LH/hCG 受体的纯化和表征以及哺乳动物 LH/hCG 受体特性的比较。

• DOI: 10.1016/0006-2952(90)90240-l
• 发表时间: 1990
• 期刊: Biochemical pharmacology
• 影响因子: 5.8
• 作者: Alpaugh,K;Indrapichate,K;Abel,JA;Rimerman,R;Wimalasena,J
• 通讯作者: Wimalasena,J