SYSTEM ANALYSIS OF PROTEIN PHOSPHORYLATION INVOLVED IN SPERM MATURATION

精子成熟过程中蛋白质磷酸化的系统分析

• 批准号: 8365483
• 负责人: Keith Roberts
• 金额: $ 5.74万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365483
• 关键词: Acrosome Reaction; Biology; Data; Drug Delivery Systems; Epididymis; Event; Female; Fertility; Funding; Grant; Informatics; Left; Modification; National Center for Research Resources; Oocytes; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Principal Investigator; Process; Proteins; Proteome; Proteomics; Research; Research Infrastructure; Resources; Source; Sperm Maturation; Staging; System; Systems Analysis; Techniques; Testis; United States National Institutes of Health; Work; contraceptive target; cost; male; reproductive; research study; sperm cell; sperm function; sperm protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Spermatozoa that leave the mammalian testis are not motile and do not have the capability to fertilize an oocyte. Sperm maturation begins in the epididymis and continues in the female reproductive tract with the process of capacitation and finally the acrosome reaction. This maturation process is unique in that sperm are transcriptionally and translationally quiescent. Sperm maturation occurs in a "closed system" where changes involve primarily modification of existing proteins. One of the biggest changes that occur is the phosphorylation states of the many sperm proteins, and these changes appear to be required for sperm function and fertility. Consequently, we propose to investigate the system wide changes which occur to the phosphoproteome of the sperm. This will be approached two ways: 1) Using the proteomic data from epididymal sperm that we have already acquired from PNNL and informatics techniques to identify which proteins have specific phosphorylation motifs that can be phosphorylated and which kinases and phosphatases are present in the sperm proteome to act on these motifs, and 2) isolate sperm at various stages of maturation and subject them to phosphoproteomics analysis to identify important maturation-related phosphorylation events. These data will be used to assemble a system analysis of the sperm phospho-proteome. These experiments will fill a significant gap in our understanding of the phosphorylation network of sperm maturation. Given the importance of kinases and phosphatases as drug targets, this work promises rapid identification of potential non-hormonal contraceptive targets in the male.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 离开哺乳动物睾丸的精子没有活力，也不具备使卵母细胞受精的能力。精子的成熟始于附睾，并在女性生殖道中继续成熟，经历获能过程，最后发生顶体反应。这种成熟过程的独特之处在于精子在转录和翻译上处于静止状态。精子成熟发生在“封闭系统”中，其中的变化主要涉及现有蛋白质的修饰。发生的最大变化之一是许多精子蛋白的磷酸化状态，这些变化似乎是精子功能和生育能力所必需的。因此，我们建议研究精子磷酸蛋白质组发生的系统范围的变化。这将通过两种方式实现：1) 使用我们从 PNNL 和信息学技术获得的附睾精子的蛋白质组数据来识别哪些蛋白质具有可以被磷酸化的特定磷酸化基序，以及精子蛋白质组中存在哪些激酶和磷酸酶来作用于这些基序，以及 2) 分离处于不同成熟阶段的精子并将其置于 磷酸化蛋白质组学分析可识别重要的成熟相关磷酸化事件。这些数据将用于对精子磷酸蛋白质组进行系统分析。这些实验将填补我们对精子成熟磷酸化网络理解的重大空白。鉴于激酶和磷酸酶作为药物靶点的重要性，这项工作有望快速识别男性潜在的非激素避孕靶点。