STRUCTURE/FUNCTION OF ZONA PELLUCIDA GLYCOPROTEINS

描述（改编自研究者摘要）：本项目 的寡糖部分的结构和功能， 小鼠透明质酸（mZP）和这种细胞外糖萼， 围绕着卵子的蛋白质与卵子表面的互补分子相互作用， 精子 先前的证据有力地表明， mZP的活性与其寡糖部分相关。 最近 申请人实验室的研究首次证明， 存在N-连接的聚乳糖胺聚糖和O-连接的聚乳糖胺聚糖， 三糖被认为在精卵中起重要作用 承认和约束。 为了评估这些生物活性， 该提案有五个具体目标：（1）纯化mZP 2 和mZP 3，然后切割和放射性标记N-连接的聚糖和O-连接的聚糖， 三糖，（2）评估放射性标记的 精卵识别和结合中的N-连接和O-连接聚糖，（3） 分离和化学表征功能性N-连接聚糖，（4）， 合成O-连接的三糖，检测其功能， 精子表面形成的机制 抗原-三糖复合物，和（5）鉴定和化学鉴定 表征小鼠精子质膜（PM）蛋白识别的 N-连接的和/或O-连接的聚糖。 完成后，这项研究将是 首次发表的关于N-连接和 O-连接寡糖。 在互补分子（即，的 ZP糖蛋白上的生物活性聚糖和精子上的互补受体 PM）在选定的实验小鼠品系中进行鉴定， 解决物种间和物种内的问题 相反配子相互作用过程中的关系。 总的来说，这些 这些研究将进一步加深我们对精卵机制的理解 相互作用和受精的小鼠，他们将提供所需的 其他植物的受精过程的调节信息 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.