Fig. 3: Overview of DiSPASO enrichment strategies.
From: Developing a new cleavable crosslinker reagent for in-cell crosslinking
A DiSPASO enrichment using a picolyl azide as a click chemistry reagent and an IMAC-based enrichment strategy to retrieve crosslinked peptides from a complex mixture. B Azide-S-S-biotin (ASSB) based enrichment using ASSB as a click chemistry reagent and biotin-streptavidin bead strategy to enrich for crosslinked peptides. The elution of bead-bound crosslinked peptides is performed using the reduction of the disulfide bond of the ASSB compound. C Simplified version of (B). The click reagent Disulfide Azide is already coupled to the beads and the click reaction is taking place directly on the beads. Elution of crosslinked peptides is performed after the click reaction and washing procedure of the beads using a reducing reagent. IUPAC names of all compounds used in this manuscript are described in Supplementary Table S4.
