STRUCTURE/FUNCTION OF ZONA PELLUCIDA GLYCOPROTEINS

描述(改编自《调查者摘要》)：本项目 重点介绍了大豆低聚糖的结构和功能 小鼠卵透明带(MZP)及其胞外糖基化 围绕着卵子与表面的互补分子相互作用 精子。先前的证据有力地表明，精子结合 MZP的活性与其低聚糖部分有关。近期 申请者实验室的研究首次证明了 N-连接的聚乳糖胺多糖和O-连接的聚乳糖胺的存在 被认为在精子-卵子中具有重要作用的三糖 认可和约束。以评估这些物质的生物活性 低聚糖，该提案有五个具体目标：(1)纯化mZP2 和mZP3，然后裂解和放射标记N-连接的多糖和O-连接的糖链 三糖，(2)评估放射性标记的功能作用 精卵识别和结合中的N-连接和O-连接的多糖，(3)到 功能性N-糖链的分离与化学表征(S)，(4)至 合成O-连接的三糖，检测其功能，并 精子表面形成的机制 抗原-三糖复合体，以及(5)鉴定和化学 识别的小鼠精子质膜(PM)蛋白的特征 N-连接和/或O-连接的葡聚糖。完成后，这项研究将成为 首次发表关于N-连接和N-连接的结构-功能多样性的报告 O-连接低聚糖。在互补分子(即 精子ZP糖蛋白和互补受体上的生物活性多糖 PM)在选定的实验小鼠品系中被鉴定，它将被 有可能解决物种间和物种内的问题 相反配子相互作用期间的关系。总而言之，这些 研究将进一步加深我们对精子-卵子机制的理解 在小鼠体内的相互作用和受精，它们将提供所需的 关于调节其他作物受精过程的信息 物种，包括人类。

DESCRIPTION (Adapted from the Investigator's Abstract): This project focuses on the structure and function of the oligosaccharide moieties of the mouse zona pellucida (mZP) and how this extracellular glycocalyx that surrounds the egg interacts with complementary molecules on the surface of spermatozoa. Previous evidence strongly suggests that the sperm binding activity of mZP is associated with its oligosaccharide portion. Recent studies in the applicant's laboratory have demonstrated for the first time the presence of N-linked polylactosaminyl glycans and an O-linked trisaccharide that are thought to have important roles in sperm-egg recognition and binding. To assess the bioactivity of these oligosaccharides, the proposal has five specific aims: (1) to purify mZP2 and mZP3, and then cleave and radiolabel the N-linked glycans and O-linked trisaccharide, (2) to assess the functional role of the radiolabeled N-linked and O-linked glycans in sperm-egg recognition and binding, (3) to isolate and chemically characterize functional N-linked glycan(s), (4) to synthesize the O-linked trisaccharide, examine its function, and the mechanism underlying the formation of the sperm surface antigen-trisaccharide complex, and (5) to identify and chemically characterize the mouse sperm plasma membrane (PM) proteins recognized by the N-linked and/or O-linked glycans. When completed, this study will be the first published report on structure-function diversity of both N-linked and O-linked oligosaccharides. After the complementary molecules (i.e., the bioactive glycans on ZP glycoproteins and complementary receptors on sperm PM) are identified in the selected strain of experimental mice, it will be possible to address the question of interspecies and intraspecies relationships during interactions of opposite gametes. Collectively, these studies will further our understanding of the mechanism of sperm-egg interaction and fertilization in the mouse, and they will provide needed information for the regulation of the fertilization processes in other species, including humans.