Regulation of Sperm Function by Protein Phosphorylation

蛋白质磷酸化调节精子功能

• 批准号: 7846469
• 负责人: SRINIVASAN VIJAYARAGHAVAN
• 金额: $ 0.69万
• 依托单位: KENT STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2009-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7846469
• 关键词: Binding; Biochemical; Biology; Cattle; Cell Cycle Progression; Cell physiology; Cyclin-Dependent Kinases; Cytosol; Development; Diagnosis; Enzymes; Epididymis; Fertilization; Genes; Germ Cells; Goals; Hamsters; Homologous Gene; Human; Immunoprecipitation; Interphase Cell; Knock-out; Knockout Mice; Knowledge; Laboratories; Male Contraceptions; Male Infertility; Mitosis; Molecular; Mus; Nuclear Protein; Nuclear Proteins; Phosphorylation; Physiological; Protein Binding; Protein Biosynthesis; Protein Dephosphorylation; Protein Isoforms; Protein Serine/Threonine Phosphatase; Proteins; Rattus; Recombinant Proteins; Regulation; Research Personnel; Role; Signal Pathway; Signal Transduction; Signaling Protein; Somatic Cell; Sperm Head; Sperm Maturation; Sperm Motility; Spermatogenesis; Testis; Time; Work; Yeasts; cell motility; enzyme activity; genetic regulatory protein; insight; male; nonhuman primate; novel; novel strategies; polypeptide; programs; protein purification; sperm cell; sperm function; zygote

DESCRIPTION (provided by applicant): A long-term goal of my laboratory is to understand the biochemical mechanisms underlying sperm motility and fertilization. In spermatozoa, where little protein synthesis occurs, protein phosphorylation is a major mechanism for regulating cellular functions. While it is known that changes in protein phosphorylation control motility initiation and fertilization, our knowledge of signaling pathways regulating these changes is limited. This proposal centers on a signaling enzyme in spermatozoa, the serine/threonine phosphatase PP1gamma2. It is a key enzyme in spermatogenesis and regulation of mature sperm function. Knowing how this enzyme works at the biochemical and molecular levels will contribute to our understanding of male gamete function. Protein purification and sequencing have enabled us to identify a number of potential PP1gamma2 binding and regulatory proteins. Immunoprecipitation and binding studies with recombinant proteins showed that two of these regulatory proteins are sds22 and protein 14-3-3 - discovered for the first time in spermatozoa. These polypeptides, which have critical functions in somatic cells, have novel roles in regulating male gamete function. In addition to changes in its binding to regulatory proteins, PP1gamma2 phosphorylation is increased during sperm maturation in the epididymis. The central hypothesis in this proposal is that the regulated action of PP1gamma2 is critical for sperm maturation, motility initiation, and other essential sperm functions. We will use biochemical and molecular approaches to characterize, at the mechanistic level, how phosphorylation and binding interactions between PP1gamma2 and its regulatory proteins relate to sperm motility and fertilizing ability. Aside from its basic importance to male gamete biology, this study has potential applications in the diagnosis and treatment of male infertility and in the development of novel approaches to male contraception.

描述（申请人提供）：我实验室的长期目标是了解精子运动和受精的生化机制。在精子中，很少发生蛋白质合成，蛋白质磷酸化是调节细胞功能的主要机制。虽然已知蛋白质磷酸化的变化控制运动起始和受精，但我们对调节这些变化的信号通路的了解有限。 这个建议集中在精子中的信号酶，丝氨酸/苏氨酸磷酸酶PP 1 γ 2。它是精子发生和调节成熟精子功能的关键酶。了解这种酶在生物化学和分子水平上的作用将有助于我们理解雄性配子的功能。蛋白质纯化和测序使我们能够确定一些潜在的PP 1 γ 2结合和调节蛋白。重组蛋白的免疫沉淀和结合研究表明，这些调节蛋白中有两个是sds 22和蛋白14-3-3 -，这是首次在精子中发现。这些在体细胞中具有关键功能的多肽在调节雄性配子功能方面具有新的作用。除了其与调节蛋白结合的变化外，PP 1 γ 2磷酸化在附睾精子成熟过程中增加。 该建议的中心假设是，PP 1 γ 2的调节作用对精子成熟、运动启动和其他基本精子功能至关重要。我们将使用生物化学和分子生物学的方法来表征，在机械水平上，如何磷酸化和PP 1 γ 2和它的调节蛋白之间的相互作用与精子活力和可移植性。除了对男性配子生物学的基本重要性外，这项研究在男性不育症的诊断和治疗以及男性避孕新方法的开发方面具有潜在的应用价值。