NMR STUDIES OF BIOLOGICALLY IMPORTANT OLIGOSACCHARIDES

具有重要生物学意义的低聚糖的核磁共振研究

• 批准号: 6498687
• 负责人: ANTHONY STEPHEN SERIANNI
• 金额: $ 22.9万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6498687
• 关键词: aminosugar; carbohydrate structure; carbon; chemical binding; combinatorial chemistry; conformation; crystallization; deuterium; glycosidases; glycosylation; method development; molecular biology information system; molecular dynamics; monosaccharides; nitrogen; nuclear magnetic resonance spectroscopy; oligosaccharides; radiotracer; sialate; stable isotope

Description: (From the applicant's abstract) Oligosaccharides are of central importance in the development and maintenance of biological systems. They are found free in solution, as constituents of glycoproteins and glycolipids on the cell surface, and have been implicated in the mechanisms of action of diverse cellular processes such as inflammation, cell-cell recognition and adhesion, protein turnover, and bacterial infection. Oligosaccharides, or molecular mimics thereof, are becoming more frequent targets in drug design for the treatment of human illness and disease. The cellular functions of oligosaccharides are intimately related to their three-dimensional shapes or conformations, and knowledge of these 3D structures, both free in solution and in the bound state, is essential to deciphering the underlying factors that control recognition between oligosaccharides and specific cellular receptors. This proposal aims to develop new probes of oligosaccharide conformation based on NMR parameters involving 13C and 2H. Major objectives are to fully establish the structural dependencies of JCH and JCC scalar couplings using experimental and computational methods, and to apply these couplings to investigate O-glycoside linkage conformation in the range of biologically important 13C-labeled di-, tri-, and tetrasaccharides, either free in solution or tethered to oligopeptides. Scalar couplings involving 15N will also be studied. Residual 13C-1H and 13C-13C dipolar couplings and residual 2H quadrupolar couplings in oriented media, and 13C chemical shift anisotropy, will be investigated as additional conformational constraints in solution. Motional properties will be investigated using molecular dynamics methods and these data compared to experimental data in order to assist in the interpretation of the latter. Investigations will then be extended to larger, biologically relevant systems including a complex high-mannose N-glycan found on CD2, engineered N-glycans on IgG, and DNA aptamer-oligosaccharide complexes.

描述：（从申请人的摘要中）寡糖是中心的 在生物系统的开发和维护中的重要性。他们是 在溶液中自由发现，作为糖蛋白和糖脂的成分 细胞表面，并与多种作用机理有关 细胞过程，例如炎症，细胞识别和粘附， 蛋白质更新和细菌感染。寡糖或分子 模仿其，在药物设计中越来越频繁地针对 治疗人类疾病和疾病。细胞函数 寡糖与它们的三维形状密切相关或 构象和对这些3D结构的知识，既可以在解决方案中自由和 在边界状态下，对破译的基本因素至关重要 寡糖和特定细胞受体之间的控制识别。 该建议旨在开发基于寡糖构象的新探针 关于涉及13C和2H的NMR参数。主要目标是完全建立 使用实验性的JCH和JCC标量耦合的结构依赖性 和计算方法，并应用这些耦合进行调查 O-糖苷链接构象在生物学上重要的范围 13C标记的DI-，三糖和四糖，可以在溶液中免费或 束缚在寡肽上。还将研究涉及15N的标量耦合。 残留的13C-1H和13C-13C偶极耦合和残留的2H四极了 定向培养基和13C化学位移各向异性的耦合将是 被研究为溶液中的其他构象约束。动作 将使用分子动力学方法和这些数据研究属性 与实验数据相比，以帮助解释 后者。然后，调查将扩展到较大的生物学相关 系统包括在CD2上找到的复杂的高甘露糖N-聚糖 IgG上的N-聚糖和DNA适体 - 寡糖复合物。