METABOLIC MONITORING OF GAG SYNTHESIS - TC3

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目是一个更广泛的综合技术项目的一部分，该项目旨在对糖胺聚糖(GAG)功能进行生化和细胞表征。GAG是形成哺乳动物细胞外基质的重要组成部分的聚合的、不同硫化的碳水化合物。蛋白质与这种基质的相互作用在引导细胞迁移、调节细胞信号和调节细胞外液和结构的物理性质方面起着重要作用。表征GAG合成和修饰的途径，并开发在体外和体内监测这些途径的方法学，对于理解功能是必不可少的。然而，这并不是一项容易的任务；GAG的硫酸盐化模式增加了聚合物的极端异质性，而且简单产品的降解和分离并不总是一种选择。理想情况下，将开发出对硫代位置敏感、甚至在体内也适用的方法。核磁共振提供了一种可能的方法。因此，正在开发将核磁共振活性同位素(15N)引入GAGS并使用二维15N-1H谱来表征硫化位置和聚合物类型的程序。此外，正在开发将信号增强到体内监测导致GAG的代谢过程所需的水平的方法。

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 a part of a more general integrated technology project directed at biochemical and cellular characterization of glycosaminoglycan (GAG) function. GAGs are polymeric, differentially sulfated, carbohydrates forming an important part of the extracellular matrix of mammalian cells. Interactions of proteins with this matrix are important in directing cell migration, modulating cell signaling and regulating the physical properties of the extracellular fluids and structures. Characterizing the pathways by which GAGs are synthesized and modified, and developing methodology by which these pathways can be monitored in vitro and in vivo, is essential to understanding function. This, however, is not an easy task; the sulfation patterns of GAGs add extreme heterogeniety to the polymers, and degradation and isolation of simple products is not always an option. Ideally, methods that are sensitive to sufation position and applicable even in vivo would be developed. Nuclear magnetic resonance (NMR) provides one possible approach. Procedures for introducing NMR active isotopes (15N) into GAGs and using two dimensional 15N-1H spectra to characterize sulfation position and polymer types are therefore being developed. Also, methodology for the enhancement of signals to the level needed for in vivo monitoring of the metabolic processes leading to GAGs is being developed.