Chapter 11: Application of HDX–MS for the Structural Characterization of Glycoproteins

Glycosylation is one of the most common and complex post-translational modifications of proteins, rendering many proteins evasive to structural analysis. The intrinsic heterogeneity and flexibility pose major challenges for most traditional structural elucidation tools. Hydrogen/deuterium exchange coupled to mass spectrometry (HDX–MS) is an increasingly widespread tool for monitoring protein dynamics, including glycoproteins and their interactions in fully glycosylated states. Recent advances have largely alleviated many of the historical challenges of applying HDX–MS to complex glycoproteins to enable the in-depth characterization of glycoproteins and even pinpoint the local structural effects of glycosylation.