Epididymis-specific proteins for male contraception

用于男性避孕的附睾特异性蛋白

• 批准号: 6442075
• 负责人: FRANK S FRENCH
• 金额: $ 3.9万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-14 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442075
• 关键词: China; carboxylic ester hydrolases; complementary DNA; contraceptives; cooperative study; endoribonucleases; epididymis; fertility; gene expression; genetic library; immunocytochemistry; male; molecular biology information system; polymerase chain reaction; protein localization; protein structure function; recombinant proteins; sperm; spermatogenesis; tissue /cell culture

DESCRIPTION (provided by applicant) This research will be performed primarily at the Shanghai Institute of Biochemistry in China as an extension of NIH grant #R37 HD04466, the parent grant for this project. The Principal and Co-Investigators of this parent grant are Frank S. French, M.D. and Susan H. Hall, PhD., the Laboratories for Reproductive Biology of the University of North Carolina at Chapel Hill. The long-term objective is to identify human epididymal targets for male contraception by investigating epididymis-specific proteins involved in sperm maturation and fertilization. The study is part of a larger contraceptive development project involving a collaboration of the Laboratories for Reproductive Biology with Professor AJ Rao at the Indian Institute of Sciences for evaluation of lead proteins in monkeys and Dr. Catherine VandeVoort at University of California, Davis Primate Center, to test the effects of immunization against epididymis secretory proteins on sperm function in monkeys. Specific Aim 1 Define expression of the human proteins corresponding to monkey Clones ESC46I-ribonuclease A and ESC615-carboxylesterase B, Specific Aim 2 Determine the functions of the human ESC461 protein Specific Aim 3 Determine the functions of ESC615 protein Specific Aim 4 Evaluate additional epididymis-specific clones for potential as male contraceptives From a macaque epididymis-specific cDNA library containing novel cDNAs related to known gene families, two are selected for in depth study. One, ESC461 is related to the eosinophil ribonucleases A2 and A3 which have antimicrobial activity. The other, SC615 is related to the carboxylesterases which may have roles in modification of sperm surface proteins. To investigate where the proteins are expressed and localized in the human epididymis and whether they bind sperm, human clones will be obtained and recombinant protein produced and used to raise antibodies in rabbits. Recombinant proteins produced in bacteria or in eukaryotes will be used to determine the ability of ESC46I to bind and hydrolyze RNA and to inhibit the growth of bacteria. The ability of SC615-carboxylesterase to catalyze the hydrolysis of a range of substrates will be determined. If time permits, studies will be initiated on 2 other clones from this library encoding lipocalin-like proteins.

描述（由申请人提供） 这项研究将主要在上海研究院进行 中国生物化学作为 NIH 拨款 #R37 HD04466 的延伸，母公司 该项目的补助金。本次母基金的主要负责人和联合研究员 是 Frank S. French 医学博士和 Susan H. Hall 博士，他们的实验室 北卡罗来纳大学教堂山分校的生殖生物学。这 长期目标是确定男性附睾的目标 通过研究精子中涉及的附睾特异性蛋白质来避孕 成熟和受精。该研究是一项更大的避孕措施的一部分 涉及实验室合作的开发项目 印度科学院 AJ Rao 教授的生殖生物学 评估猴子体内的先导蛋白，Catherine VandeVoort 博士在 加州大学戴维斯分校灵长类动物中心，测试效果 附睾分泌蛋白免疫对精子功能的影响 猴子。 具体目标 1 定义与猴子相对应的人类蛋白的表达 克隆 ESC46I-核糖核酸酶 A 和 ESC615-羧酸酯酶 B，具体目标 2 确定人 ESC461 蛋白的功能 具体目标 3 确定 ESC615 蛋白的功能 具体目标 4 评估其他 附睾特异性克隆具有作为男性避孕药的潜力 来自猕猴附睾特异性 cDNA 文库，其中包含相关的新型 cDNA 在已知的基因家族中，选择其中两个进行深入研究。一、ESC461是 与具有抗菌作用的嗜酸性粒细胞核糖核酸酶 A2 和 A3 有关 活动。另一方面，SC615 与羧酸酯酶有关，可能具有 精子表面蛋白修饰中的作用。去调查哪里 蛋白质在人类附睾中表达和定位，以及它们是否 结合精子，获得人类克隆并产生重组蛋白 用于在兔子体内产生抗体。细菌产生的重组蛋白 或在真核生物中将用于确定 ESC46I 结合和的能力 水解RNA并抑制细菌生长。的能力 SC615-羧酸酯酶催化一系列底物的水解 被确定。如果时间允许，将开始对另外 2 个克隆进行研究 来自该编码脂质运载蛋白样蛋白的文库。