Fig. 2: Establishment of a streamlined chemoproteomic workflow for PBSP profiling. | Nature Communications

Fig. 2: Establishment of a streamlined chemoproteomic workflow for PBSP profiling.

From: Spatiotemporal profiling of modification-specific proteome secretion uncovers an itaconation-activated tyrosine kinase

Fig. 2

a Schematic of the FISAP platform. A custom C18 tip, pre-loaded with streptavidin beads, was first treated with sodium dodecyl sulfate (SDS) to inactivate the C18. ITalk-labeled proteins extracted from the culture medium were click-reacted with azide-biotin and then incubated with the streptavidin beads for enrichment. After removing unlabeled proteins by centrifugation, the C18 was re-activated, and on-bead trypsin digestion was performed. The released peptides were desalted directly by the C18, followed by elution for DIA-based LC-MS/MS analysis. Created in BioRender. Chu, l. (2025) https://BioRender.com/e55b8pm. b Schematic of the DIA-based LC-MS/MS analysis. c Correlation of protein intensities quantified by the PBSP workflow across three biological replicates. d Volcano plot showing the comparative enrichment of proteins between ITalk labeling group and control group by PBSP. Itaconated secreted proteins are highlighted in red. p-values were determined by two-sided Student t-tests.

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