A key challenge for glycoscience is in the discovery of glycan-mediated binding interactions. Glycans mediate many physiologically critical recognition events, however glycan-protein interactions are typically low-affinity, often exhibiting micromolar or even millimolar equilibrium dissociation constants. Notwithstanding these low affinities, glycan recognition events can be highly specific, often through multivalent presentation of glycans and/or their ligands on cell surfaces. Typical biochemical methods for interaction discovery break apart cells, resulting in disruption of and scrambling of glycan-protein interactions. We developed methods to incorporate the diazirine photocrosslinker onto GlcNAc residues of both intracellular and extracellular glycans. Applying this metabolic oligosaccharide engineering appoach, it is possible to introduce a covalent crosslink between glycans and their binding partners while in the normal cellular context. Once this crosslink is formed, cells can be lysed and the covalent complexes analyzed by common methods such as immunoblot and mass spectrometry-based proteomics. I will describe recent improvements to GlcNAc crosslinking methodology and application to various glycan-dependent binding interactions.

Use of photocrosslinking N-acetylglucosamine (GlcNAc) to define glycan-dependent binding interactions

Where thousands of chemistry professionals meet to share ideas and advance scientific and technical knowledge. Spring 2023 abstracts, posters, and presentations will launch at the end of March.

ACS Spring 2023

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.