Fig. 2: Development of a highly selective proximity proteomics approach for temporal membrane-associated and cytosolic interactome. | Nature Communications

Fig. 2: Development of a highly selective proximity proteomics approach for temporal membrane-associated and cytosolic interactome.

From: Spatiotemporal profiling of cytosolic signaling complexes in living cells by selective proximity proteomics

Fig. 2

a Scheme of BP5-based proximity proteomic analysis of adapter protein interactome upon EGF stimulation in living cells. b, c Volcano plots of GRB2-interacting proteins in HeLa cells with stably expressed APEX2-FLAG-GRB2 quantified by BP5- and BP1-based proximity proteomics (n = 3 independent biological experiments). Significantly changed proteins upon 2 min EGF stimulation are highlighted in red (FDR < 0.05 and S0 = 0.5). Relevant proteins that are significant in BP5 experiment but not in BP1 experiment are marked in green. d Interaction map and function annotation of significant GRB2-associated proteins identified by BP5- and BP1-proximity proteomics. The colored circles indicate the functional classification. Known GRB2- associated proteins were labeled in red. e Comparison of BP5 and BP1 labeling selectivity as indicated by both known GRB2-associated proteins and cytosolic proteins with GO annotation of cytosol identified from the same pull-down sample (insert). The average ratio of LFQ intensity (BP5/BP1) from three replicates were plotted. Source data are provided as a Source Data file.

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