NEW STRATEGIES TO DETERMINE STRUCTURE-FUNCTION RELATIONSHIPS OF HS CO-RECEPTORS

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。 HS通过单独与配体相互作用或同时与配体和受体相互作用而发挥辅助受体的作用。这些相互作用模式被各种生长因子、形态因子和细胞外基质(ECM)蛋白所共享。使用低聚糖的生物化学研究已经证实，一般而言，HS需要不同的结构来与不同的蛋白质相互作用。然而，HS在细胞环境中作为辅助受体的结构-功能关系要复杂得多。这在一定程度上是因为，天然HS比生化分析中使用的低聚糖大得多，可以通过不同的结合位点同时与配体及其受体相互作用，还可能含有多个结合位点的副本。在这里，我们的目标是开发基于细胞的分析，以扩展和验证细胞环境中的生化发现，以更好地了解HS的生物学功能。特别是，我们正在利用我们产生的系列HS突变小鼠肺内皮细胞(MLEC)作为模型系统来开发基于细胞的分析来确定HS共受体功能的一般作用和结构-功能关系。

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. HS functions as a co-receptor by interacting with ligands alone or with both ligands and receptors. These modes of interaction are shared by a variety of growth factors, morphogens and extracellular matrix (ECM) proteins. Biochemical studies using oligosaccharides have established that, in general, HS requires distinct structures to interact with different proteins. However, the structure-function relationship of HS in its role as a co-receptor in a cellular context is far more complex. This in part results from the fact that natural HS is much larger than the oligosaccharides used in biochemical analyses, may interact simultaneously with both ligand and its receptor via distinct binding sites, and may also harbor multiple copies of the binding sites. Here we aim to develop cell-based assays to extend and to validate biochemical findings in a cellular context for a better understanding of the biological functions of HS. In particular, we are utilizing our generated serial HS mutant mouse lung endothelial cells (MLEC) as the model system to develop cell-based assays to determine the general role and the structure-function relationship of HS co-receptor functions.