HYALURONAN-PROTEIN INTERACTIONS USING HYDROGEN/DEUTERIUM EXCHANGE MS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。蛋白质-透明质酸(HA)相互作用在细胞外基质组装、细胞黏附、肿瘤转移和免疫反应等多种生物现象中具有重要意义。因此，研究蛋白质-HA相互作用的工具对于进一步了解它们的功能是必不可少的。氢/氢(H/D)酰胺交换质谱学为监测蛋白质骨架表面暴露的酰胺氢的自由交换提供了一种有效的方法。这些酰胺氢是不稳定的，当暴露在氧化重氢中时，可以被重氢取代，导致每个酰胺增加1道尔顿。为了研究蛋白质-HA的结合，在没有和存在HA寡糖配体的情况下，分析了蛋白质中的氚掺入水平。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Protein-hyaluronan (HA) interactions are of profound importance in diverse biological phenomena such as extracellular matrix assembly, cell adhesion, tumor metastasis and the immune response. Therefore, tools for studying protein-HA interactions are essential to gain further insight into their function. Hydrogen/deuterium (H/D) amide-exchange mass spectrometry provides a powerful method for monitoring the free exchange of surface exposed amide hydrogens on a protein backbone. These amide hydrogens are labile and can be replaced by deuterons when exposed deuterium oxide results in an increase in 1 Dalton for each amide. To investigate protein-HA binding, the level of deuterium incorporation in the protein is analyzed in the absence and presence of the HA oligosaccharide ligand.