UC Irvine

The following document describes the synthesis and preliminary testing of three novel CID-cleavable crosslinking agents for the study of protein-protein interactions. These crosslinkers contain a sulfoxide group that can be cleaved inside of the mass spectrometer at a lower energy than the peptide backbone, allowing for a series of tandem experiments to identify peptide fragments. Three crosslinkers of varying length containing a lysine-reactive N-hydroxysuccinimide ester and a photoreactive diazirine ring were developed. One crosslinker was functionalized with an alkyne to allow for affinity purification with biotin. Two of the crosslinkers have been found to successfully form crosslinks within synthetic peptides and bovine serum albumin, and experiments are currently ongoing with the third crosslinking agent.