Epididymis-specific proteins for male contraception

描述(由申请人提供) 这项研究将主要在上海经济技术学院进行 中国生物化学作为美国国立卫生研究院#R37HD04466拨款的延伸 为这个项目拨款。这项家长基金的主要负责人和共同调查员 弗兰克·S·弗兰奇医学博士和苏珊·H·霍尔博士， 北卡罗来纳大学教堂山分校生殖生物学。这个 长期目标是为男性识别人类附睾靶 通过研究精子中涉及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药的一部分 涉及实验室合作的开发项目 与印度科学院AJ·拉奥教授合著《生殖生物学》 用于评估猴子体内的铅蛋白，凯瑟琳·范德沃特博士在 加州大学戴维斯灵长类中心，以测试 附睾分泌蛋白免疫对小鼠精子功能的影响 猴子。 特定目的1定义与猴子相对应的人蛋白的表达 ESC46I-核糖核酸酶A和ESC615-羧酸酯酶B克隆，特异靶2 测定人ESC461蛋白的功能特异靶3测定 ESC615蛋白特异性目标4的功能评价 附睾特异性克隆有可能作为男性避孕药 从一个猕猴附睾特异性cDNA文库中获得 对于已知的基因家族，选择了两个进行深入研究。其一，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。另一个是SC615，与羧酸酯酶有关，它可能具有 精子表面蛋白修饰中的作用。为了调查在哪里 蛋白质在人类附睾中表达和定位，以及它们是否 结合精子，将获得人类克隆并生产重组蛋白和 用来提高兔子的抗体。细菌中产生的重组蛋白 或在真核细胞中，将被用来确定ESC46I结合和 能降解RNA，抑制细菌的生长。的能力 SC615-羧酸酯酶催化一系列底物的水解会 要下定决心。如果时间允许，将启动对另外两个克隆的研究 从这个编码Lipocalin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.