We have developed photocrosslinking sugar technology that can be used to identify glycan-dependent interactions.
We synthesize unnatural analogs of common monosaccharides. The analogs contain the diazirine photocrosslinking functional group. This small, three-membered ring can be activated by UV (~350 nm) irradiation to generate a highly reactive carbene intermediate. The carbene will rapidly insert into nearby C-H, O-H, or N-H bonds, resulting in a covalent crosslink between the sugar and neighboring molecules.
These crosslinked complexes can be purified and/or analyzed by immunoblot or mass spectrometry. Photocrosslinking sugars can be used in purified systems, or can be incorporated into the glycoconjugates of living cells, enabling crosslinking of glycan-dependent or -associated interactions in their native, cellular context.