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Abstract
Glycosaminoglycans (GAGs) are linear, highly negatively charged polysaccharides expressed by all animal cells. Through interactions with many proteins, GAGs act as regulatory molecules in many physiological and pathological processes. Travelling Wave Ion Mobility Spectrometry (TWIMS), a gas phase biophysical technique that separates ions based on their size, shape and charge is a valuable tool in the structural characterization of biomolecules based on measurement of rotationally average collision cross sections (CCSs). In this dissertation, we explore the application of native mass spectrometry (native MS) coupled with TWIMS to investigate different protein-GAG binding systems. ATIII and Arixtra, a well-studied protein-GAG system with a high binding specificity, was examined to establish the capabilities of this approach. More complicated and less well characterized systems were studied, including FGF1 and Robo1 with heparin-like GAGs. Our results demonstrate the potential for applying native MS-TWIMS to achieve a molecular level understanding of the nature of these protein-GAG interactions.