HYALURONAN-PROTEIN INTERACTIONS USING HYDROGEN/DEUTERIUM EXCHANGE MS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。蛋白-透明质酸（HA）相互作用在细胞外基质组装、细胞粘附、肿瘤转移和免疫应答等多种生物现象中具有重要意义。因此，研究蛋白质-血凝素相互作用的工具对于进一步了解其功能至关重要。氢/氘（H/D）酰胺交换质谱法为监测蛋白质主链表面暴露的酰胺氢的自由交换提供了一种强大的方法。这些酰胺氢是不稳定的，当暴露在氧化氘中导致每个酰胺增加1道尔顿时，它们可以被氘核取代。为了研究蛋白与血凝素的结合，分析了在缺乏和存在血凝素寡糖配体的情况下，蛋白质中氘的掺入水平。

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.