赖氨酸乙酰化修饰与精子成熟调控

项目名称： 赖氨酸乙酰化修饰与精子成熟调控

项目编号： No.31300959

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 于合国

作者单位： 上海市计划生育科学研究所

项目金额： 25万元

中文摘要： 精子从发生到成熟经历了一系列的修饰作用，其中精子蛋白翻译后修饰已被证实与精子运动、获能、结合透明带及受精能力相关，而赖氨酸乙酰化修饰是近年来翻译后修饰研究的热点，但是其在精子成熟中的作用及意义鲜见报道。在前期工作中，我们成功制备了高特异性的赖氨酸乙酰化单克隆抗体，发现小鼠附睾精子从附睾头到附睾尾乙酰化修饰变化明显，并且我们在人精子上鉴定出大量乙酰化修饰蛋白，而这些蛋白与精子的代谢和信号传导等密切相关，并且乙酰化酶和去乙酰化酶抑制剂对体外培养精子的运动有明显的抑制作用，提示乙酰化修饰可能在精子成熟过程发挥重要的调控作用。本项目拟在上述工作基础上，筛选出精子成熟过程中通过乙酰化修饰发挥重要调控作用的靶蛋白，探讨其乙酰化修饰的具体功能及其在精子成熟中的作用机理，以期加深我们对精子成熟调控机制的了解；并通过收集和检测正常和不育人群的各类精子样本，初步分析赖氨酸乙酰化修饰在临床诊治中的应用前景。

中文关键词： 精子；乙酰化；单克隆抗体；获能；乙酰化转移酶

英文摘要： From its generation to maturation, spermatozoon undergoes series of modifications, among which post translational modification of sperm proteins,has been proved to be involved in sperm motility, capacitation, sperm-zona binding and sperm fertilizing capacity. Lysine acetylation, as a hotspot in post translational modification researches,has not been reported regarding its roles and significance in sperm maturation.We have raised highly specific monoclonal antibodies that recognize acetylated Lysine . With these antibodies,we observed dramatic changes of lysine acetylation profiles of mice sperms from caput epididymis to cauda epididymis. Besides, we also determined the lysine acetylayion profiles of a large array of human sperm proteins.These proteins with lysine acetylayion have been correlated with sperm metabolic regulation and cell signaling in bioformatic analysis. In addition, incubation of sperms with inhibitors of acetyltransferases and deacetyltransferases resulted in sperm motility inhibition in vitro, which suggested that lysine acetylation and deacetylayion should play key roles in the regulation of sperm maturation. Therefore, with an aim to further understand the regulatory mechanisms of sperm maturation, we plan to identify key proteins that regulate sperm maturation through lysine acetylation mod

英文关键词： sperm；acetylation；monoclonal antibody；capacitation；acetyltransferase