A MOLECULAR APPROACH TO GONADOTROPIN RECEPTOR REGULATION

促性腺激素受体调节的分子方法

• 批准号: 3320172
• 负责人: JAY wimalasena WIMALASENA
• 金额: $ 9.9万
• 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/3320172
• 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受体的程序