Fig. 5: Comparison of DiSPASO and DSBSO using Cas9 as a model protein, as well as enrichment performance of DiSPASO using picolyl azide as enrichment tag.
From: Developing a new cleavable crosslinker reagent for in-cell crosslinking
A Number of identified unique crosslinks (unique residue pairs) in a triplicate experiment of Cas9 crosslinked with either DiSPASO or DSBSO in different injection amounts (500 and 200 ng). Blue indicates non-enriched crosslinked peptides, yellow enriched crosslinked peptides from Cas9 DiSPASO experiments. The green color represents Cas9 crosslinking results using DSBSO. The mean value of residue pairs is plotted in the middle of the bar. The number of technical replicates is indicated as separate black dots on top of the bar (n = 3). The standard deviation was estimated as average distance from each data point to the sample mean. B Overlap of identified crosslinks after click reaction and enrichment using picolyl azide as click chemistry reagent. After click reaction and enrichment using picolyl azide the numbers of crosslinks show an overlap of 49% with 21% unique to the enriched crosslinked sample. Left bottom of B: Overlap of technical replicates of non-enriched Cas9 links. Right bottom of B: Overlap of technical replicates of enriched Cas9 links.
