HYALURONAN-PROTEIN INTERACTIONS USING HYDROGEN/DEUTERIUM EXCHANGE MS

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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.