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所进行 中国的生物化学作为NIH赠款＃R37 HD04466的扩展 授予这个项目。该父母赠款的主要和共同研究员 M.D. Frank S. French和Susan H. Hall博士是实验室 北卡罗来纳大学教堂山的生殖生物学。这 长期目标是确定男性的人类附子靶标 通过研究参与精子的附睾特异性蛋白来避孕 成熟和受精。该研究是较大避孕药的一部分 开发项目涉及实验室的合作 印度科学研究所的AJ Rao教授的生殖生物学 用于评估猴子中的铅蛋白和凯瑟琳·范德沃特博士 加利福尼亚大学戴维斯灵长国中心，以测试 针对附睾分泌蛋白的免疫对精子功能的免疫 猴子。 特定目标1定义与猴子相对应的人蛋白的表达 克隆ESC46I-核糖酶A和ESC615-羧基酯酶B，特定目标2 确定人ESC461蛋白特异性目标的功能3确定 ESC615蛋白特异性目标的功能4评估了其他 代替男性避孕药的附睾特异性克隆 来自猕猴的附加膜特异性cDNA库，其中包含新型cDNA相关的 对于已知的基因家族，选择了两个进行深入研究。一个，Esc461是 与含有抗菌剂 活动。另一个SC615与可能具有的羧酸酯酶有关 在修饰精子表面蛋白质中的作用。调查在哪里 蛋白质被表达并局部在人类附子症中，以及它们是否存在 结合精子，将获得人类克隆并产生重组蛋白，并 用于饲养兔子中的抗体。在细菌中产生的重组蛋白 或在真核生物中，将用于确定ESC46i结合和 水解RNA并抑制细菌的生长。能力 SC615-羧基酯酶催化一系列底物的水解将 可以确定。如果时间允许，将在其他两个克隆上开始研究 从这个编码Lipocalin样蛋白的库中。