NMR METHODOLOGY FOR CHARACTERIZING CARBOHYDRATE-PROTEIN INTERACTIONS (TB1)

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目是针对糖胺聚糖-蛋白质复合物的结构表征的更一般的综合技术项目的一部分。 一般来说，碳水化合物-蛋白质复合物的形成在涉及细胞与其环境相互作用的各种过程中是重要的。这些过程包括生理过程，如细胞分化、细胞聚集和细胞信号传导;它们还包括疾病过程，如病毒感染、恶性肿瘤和不必要的炎症。开发药物以缓和自然过程和抑制疾病过程始于寡糖-蛋白质相互作用的精确结构模型。由于寡糖-蛋白质复合物中缺少短距离NOE接触，这些模型很难通过传统的核磁共振（NMR）方法得到。我们正在开发一些实验和分析工具，当传统的NMR方法失败时，可以检索结构信息。这个特殊的项目结合了Moremen实验室开发的新同位素标记策略，新的分配方法和新类型的NMR可观察的结构数据。 靶系统包括糖基化蛋白质和高阶聚集体，糖基化蛋白质难以以适于使用NMR策略的形式表达，所述NMR策略依赖于用多种NMR活性同位素的均匀标记，高阶聚集体包括趋化因子-糖胺聚糖（GAG）聚集体，其中分辨率和灵敏度成为问题。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project is part of a more general integrated technology project directed at the structural characterization of glycoaminoglycan-protein complexes. The formation of carbohydrate-protein complexes, in general, is important in a variety of processes involving the interaction of a cell with its environment. These processes include physiological processes, such as cell differentiation, cell aggregation, and cell signaling; they also include disease processes, such as viral infection, malignancy, and unwanted inflammation. The development of drugs to moderate natural processes and inhibit disease processes begins with accurate structural models for oligosaccharide-protein interactions. These models have been difficult to get by traditional Nuclear Magnetic Resonance (NMR) approaches because of the dearth of short-distance NOE contacts in oligosaccharide-protein complexes. We are developing a number of experiments and analysis tools that allow retrieval of structural information when traditional NMR approaches fail. This particular project combines new isotopic labeling strategies developed in the Moremen laboratory with new methods of assignment, and new types of NMR-observable structural data. The target systems include both glycosylated proteins that are difficult to express in forms amenable to use of NMR strategies that are dependent on uniform labeling with multiple NMR-active isotopes, and high order aggregates including chemokine-glycosaminoglycan (GAG) aggregates, where resolution and sensitivity become a problem.