中央研究院 生物化學研究所

Glycan-based cross-linking mass spectrometry (XL-MS) has recently emerged as a promising methodology capable of probing glycan-dependent interactions in complex biological systems. By harnessing metabolic glycoengineering (MGE) to introduce biologically inert azide groups within specific monosaccharides in a given oligosaccharide on secreted and membrane glycoproteins, glycans can be chemically ligated to neighbouring protein interactors using hetero-bifunctional crosslinkers. While glycan XL-MS has so far been applied to the cell surface interactome via NHS-based linkers, expansion of this technology through the development of bespoke azide-targeting linkers with different chemical properties for targeted analysis has yet to be explored. Here, we describe the generation of a novel hetero-bifunctional photoreactive linker containing DBCO and diazirine functional groups, the optimisation of MGE through cell culture manipulation and ion mobility MS (IM-MS) quantification, which in combination we applied to uncover receptor and lectin binding to coronavirus glycoprotein spikes via glycan XL-MS.