Regulation of Sperm Function by Protein Phosphorylation

蛋白质磷酸化调节精子功能

基本信息

批准号：
7659309
负责人：
SRINIVASAN VIJAYARAGHAVAN
金额：
$ 1.9万
依托单位：
KENT STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-06-01 至 2009-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659309
关键词：
Binding Biochemical Biology Bos taurus 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 Localized Male Contraceptions Male Infertility Mitosis Molecular Mus Nuclear Protein Nuclear Proteins Numbers 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 knockout gene male nonhuman primate novel novel strategies polypeptide programs protein purification sperm cell sperm function zygote

项目摘要

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（由申请人提供）：我的实验室的一个长期目标是了解精子运动和施肥的生化机制。在几乎没有蛋白质合成的精子中，蛋白质磷酸化是调节细胞功能的主要机制。虽然众所周知，蛋白质磷酸化控制运动的起始和受精的变化，但我们对调节这些变化的信号通路的了解是有限的。该建议集中在精子中的信号传导酶，丝氨酸/苏氨酸磷酸酶PP1GAMMA2。它是成熟精子功能的精子发生和调节的关键酶。了解这种酶在生化和分子水平上的工作方式将有助于我们对男配子功能的理解。蛋白质纯化和测序使我们能够识别许多潜在的PP1GAMMA2结合和调节蛋白。与重组蛋白的免疫沉淀和结合研究表明，这些调节蛋白中的两种是SDS22和蛋白14-3-3-在精子中首次发现。这些在体细胞中具有关键功能的多肽在调节男配子功能中具有新的作用。除了其与调节蛋白的结合变化外，在附睾精子成熟期间，PP1GAMMA2磷酸化增加了。该提议中的中心假设是，PP1GAMMA2的调节作用对于精子成熟，运动启动和其他必要的精子功能至关重要。我们将使用生化和分子方法在机械水平上表征PP1GAMMA2及其调节蛋白之间的磷酸化和结合相互作用如何与精子运动和受精能力有关。除了其对男配子生物学的基本重要性外，这项研究在诊断和治疗男性不育症以及开发新型男性避孕方法方面还具有潜在的应用。