STRUCTURE/FUNCTION OF ZONA PELLUCIDA GLYCOPROTEINS

透明带糖蛋白的结构/功能

• 批准号: 2673966
• 负责人: DAULAT R TULSIANI
• 金额: $ 16.77万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2673966
• 关键词: cell cell interaction; egg /ovum; fertilization; high performance liquid chromatography; laboratory mouse; laboratory rabbit; oligosaccharides; protein structure function; sperm; surface antigens; zona pellucida glycoproteins

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.

描述（改编自研究者摘要）：该项目 重点研究寡糖部分的结构和功能 小鼠透明带 (mZP) 以及这种细胞外糖萼如何 围绕鸡蛋与表面上的互补分子相互作用 精子。 先前的证据强烈表明精子结合 mZP 的活性与其寡糖部分相关。 最近的 申请人实验室的研究首次证明 N-连接的聚乳糖胺聚糖和 O-连接的存在 三糖被认为在精卵中具有重要作用 识别和约束。 为了评估这些物质的生物活性 寡糖，该提案有五个具体目标：（1）纯化mZP2 和 mZP3，然后切割并放射性标记 N-连接聚糖和 O-连接聚糖 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 连接聚糖，(3) 分离和化学表征功能性 N 连接聚糖，(4) 合成O-连接三糖，检查其功能，并 精子表面形成的机制 抗原-三糖复合物，以及(5)鉴定和化学 表征小鼠精子质膜 (PM) 蛋白质的识别 N-连接和/或O-连接聚糖。 完成后，这项研究将是 首次发表关于 N 联结构和功能多样性的报告 O-连接寡糖。 在互补分子（即， 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被鉴定出来，这将是 可能解决种间和种内问题 异配子相互作用过程中的关系。 总的来说，这些 研究将进一步加深我们对精卵机制的理解 小鼠体内的相互作用和受精，它们将提供所需的 其他受精过程调节的信息 物种，包括人类。