2015 Volume 27 Issue 156 Pages J1-J7
Photocrosslinking approaches are an important strategy to covalently capture biomolecular interactions. Covalent capture by photocrosslinking is particularly powerful for glycan-dependent interactions because these binding events are typically low-affinity and unable to withstand common purification protocols. Implementation of photocrosslinking methods is based on the incorporation of a photoactivatable crosslinking functional group onto glycan-containing molecules. Upon activation with UV radiation, the photocrosslinking group is converted to a reactive species, which forms a covalent bond with neighboring molecules. Resulting crosslinked complexes can be analyzed by immunological and mass spectrometry techniques to identify glycan-dependent binding partners. In some cases, photocrosslinker incorporation can be achieved in living cells, enabling detection of glycan-dependent interactions in their native context. In this mini-review, we describe new sugar-based photocrosslinking reagents and illustrate how they have been used to photochemically characterize glycan-dependent interactions.