Fig. 1: Validation of the PG-DA probe in the mouse brain proteome.

A Chemical structures of the palmitic acid-diazarine-alkyne (PA-DA) and 1-palmitoyl-glycerol-diazarine-alkyne (PG-DA) probes used in this study. B A representative fluorescence gel from an in-gel chemoproteomics experiment showing the UV-dependent photocrosslinking of the PG-DA probe (500 μM, 6 mins of UV exposure) to proteins from the membrane (memb) and soluble (sol) proteomes prepared from the mouse brain. Coomassie staining was used to ensure equal loading in this experiment. This experiment was done three times with reproducible results each time. C A LC-MS/MS based chemoproteomics experiment showing enrichment ratio (heavy:light; H:L) of the total proteins identified from the UV-dependent photocrosslinking of the PG-DA probe (500 μM, 6 mins of UV exposure) from the membrane and soluble proteomes prepared from the mouse brain. Each data point represents the mean of the enrichment ratio obtained for the respective protein from two or three biological replicate for a particular proteomic fraction, based on the defined filtering criteria for this proteomics experiment. The horizontal dotted line denotes an enrichment ratio ≥ 3, and proteins having an enrichment ratio above this threshold were considered enriched by the PG-DA probe, and taken forward for subsequent analysis. Complete details for all the proteins can be found in Supplementary Data 1. D–F Categorization of proteins enriched by the PG-DA probe from the mouse brain proteome based on the Panther database categorization into different: (D) protein classes; (E) biological processes involved in; and (F) known molecular functions.