人睾丸蛋白质表达谱的构建及精子发生相关蛋白质组学研究

发布时间：2018-10-09 11:57

【摘要】：睾丸是重要的男性生殖腺，其主要作用是产生精子和雄性激素。在整体水平上对人睾丸的蛋白质表达谱进行分析，将为阐明精子发生以及男性生殖调控的分子机理提供重要信息。基于这一目的，本研究利用蛋白质组学的技术手段研究了正常人睾丸组织的蛋白组成并建立了人睾丸蛋白质的参考数据库。我们利用双向凝胶电泳对正常人睾丸蛋白进行了分离，在硝酸银染色后将所有可见的蛋白点切取下来，经胰蛋白酶消化以后用MALDI-TOF质谱仪进行鉴定。成功鉴定的979个蛋白点对应于541种蛋白质。这些蛋白质信息依照WORLD-2DPAGE国际双向电泳数据库联盟的标准构建了可供全球共享的人睾丸蛋白质的参考数据库(http://reprod.njmu.edu.cn/cgi-bin/2d/2d.cgi)。利用生物信息学分析我们对每一种蛋白质的分子功能、参与的生物学过程等进行了详细注释，发现约占80％的蛋白质参与了代谢过程。依据Kegg数据库和BioCarta数据库收录的代谢通路我们分析了人睾丸组织特征性的代谢途径，在两个不同数据库的分析结果中均具有显著意义的代谢通路包括糖酵解通路和精氨酸的代谢通路，此外、在睾丸组织中还存在着由泛素-蛋白酶体系统介导的蛋白质代谢等20余条特征性的代谢通路。除参与了代谢过程以外，人睾丸组织中蛋白质还具有一系列其它重要功能，如结合蛋白质和核苷酸、具有水解酶和氧化还原酶的活性、结合离子等。利用Pathway Assist软件，，我们构建了人睾丸蛋白质的分子调控网络，从中可以看出，在人睾丸组织中众多蛋白质共同参与调控的主要有细
[Abstract]:The testis are important male gonads that produce sperm and testosterone. The analysis of the protein expression profile of human testis at the whole level will provide important information for elucidating the molecular mechanism of spermatogenesis and male reproductive regulation. For this purpose, the proteomics technique was used to study the protein composition of normal human testis and the reference database of human testis protein was established. The normal human testicular proteins were separated by two-dimensional gel electrophoresis. After silver nitrate staining, all the visible protein spots were cut off, then digested by trypsin and identified by MALDI-TOF mass spectrometry. The 979 protein spots identified successfully corresponded to 541 proteins. According to the standard of WORLD-2DPAGE, the reference database of human testicular protein (http://reprod.njmu.edu.cn/cgi-bin/2d/2d.cgi) is constructed. Using bioinformatics to analyze the molecular functions and biological processes of each protein, we found that about 80% of the proteins were involved in the metabolic process. Based on the metabolic pathways recorded in Kegg and BioCarta databases, we analyzed the characteristic metabolic pathways of human testis. Significant metabolic pathways were found in two different databases, including glycolytic pathway and arginine pathway. There are more than 20 characteristic metabolic pathways such as protein metabolism mediated by ubiquitin-proteasome system in testis. In addition to participating in the metabolic process, proteins in human testis also have a series of other important functions, such as binding proteins and nucleotides, hydrolase and oxidoreductase activities, binding ions and so on. Using Pathway Assist software, we constructed the molecular regulatory network of human testicular protein, from which we can see that in human testicular tissue, many proteins are mainly involved in the regulation.
【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R321

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