Epididymis-specific proteins for male contraception

描述（由申请人提供） 这项研究将主要在上海研究所进行。 中国生物化学作为NIH资助#R37 HD 04466的延伸， 资助这个项目。本母基金的主要研究者和合作研究者 弗兰克S。法语，医学博士苏珊H。Hall博士，实验室为 查佩尔山的北卡罗来纳州大学生殖生物学。的 长期目标是确定男性的人类附睾靶点 通过研究精子中的附睾特异性蛋白来避孕 成熟和受精。这项研究是一项更大规模的避孕计划的一部分。 涉及实验室合作的开发项目， 生殖生物学与AJ Rao教授在印度科学研究所 用于评估猴子中的铅蛋白质，凯瑟琳·范德沃特博士在 加州大学戴维斯灵长类动物中心，以测试 附睾分泌蛋白免疫对精子功能影响 猴子 具体目标1定义与猴蛋白质相对应的人蛋白质的表达 克隆ESC 46 I-核糖核酸酶A和ESC 615-羧酸酯酶B，特异性目标2 确定人ESC 461蛋白的功能特异性目的3确定 ESC 615蛋白的功能特异性目的4 附睾特异性克隆作为男性避孕药的潜力 从猕猴附睾特异性cDNA文库中筛选出与附睾相关的新cDNA， 从已知的基因家族中，选择了两个进行深入研究。ESC 461是 与嗜酸性粒细胞核糖核酸酶A2和A3相关， 活动另一个是SC 615，它与羧酸酯酶有关， 精子表面蛋白的修饰作用。去调查 蛋白质在人附睾中表达和定位，以及它们是否 结合精子，将获得人类克隆并产生重组蛋白， 用来在兔子体内产生抗体细菌中产生的重组蛋白 或在真核生物中的表达将用于确定ESC 46 I结合和 水解RNA并抑制细菌生长。的能力 SC 615-羧酸酯酶催化一系列底物的水解， 被确定。如果时间允许，将对另外2个克隆启动研究 从这个编码脂质运载蛋白样蛋白的文库中。

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.