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Here, we present a paired set of PLP workflows that simultaneously track neighborhood changes for oncogenic epidermal growth factor receptor (EGFR) with temporal resolution, both outside and inside of cells. We achieved this by augmenting the multiscale PLP workflow we call MultiMap, where three photo-probes with different labeling ranges were photo-activated by one photocatalyst, Eosin Y. By anchoring Eosin Y extracellularly and intracellularly on EGFR, we captured hundreds of proteins on both sides of the cell membrane that change in proximity to EGFR upon EGF activation. Neighbors engaged with EGFR within minutes to over an hour, reflecting dynamic interactomes during early, mid- and late-signaling including phosphorylation, internalization, degradation and transcriptional regulation. This rapid \u201cphotographic\u201d labeling approach provides snapshots of signaling neighborhoods, revealing their dynamic nature, and potential for drug targeting.<\/jats:p>","DOI":"10.1101\/2024.12.21.629933","type":"posted-content","created":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T03:06:12Z","timestamp":1735009572000},"source":"Crossref","is-referenced-by-count":1,"title":["Spatiotemporal photocatalytic proximity labeling proteomics reveals ligand-activated extracellular and intracellular EGFR 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