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Temporal photoproximity labeling of ligand-activated EGFR neighborhoods using MultiMap

ABSTRACT: Photoproximity labeling proteomics (PLP) methods have recently shown that cell surface receptors can form lateral interactome networks. Here, we present a paired set of PLP workflows that dynamically track neighborhood changes for oncogenic epidermal growth factor receptor (EGFR) over time, both outside and inside of cells. We achieved this by augmenting the multiscale PLP workflow we call MultiMap, where three photoprobes with different labeling ranges were photoactivated by one photocatalyst, eosin Y, anchored extracellularly and intracellularly on EGFR. We identified hundreds of neighboring proteins that changed within minutes to over 1 h after the addition of EGF. These neighborhoods reveal dynamic interactomes during early, middle and late signaling that drive phosphorylation, internalization, degradation and transcriptional regulation. This rapid ‘molecular photographic’ labeling approach provides snapshots of signaling neighborhoods, revealing their dynamic nature and potential for drug targeting.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Zhi Lin

LAB HEAD: Zhi Lin

PROVIDER: PXD059180 | Pride | 2025-11-18

REPOSITORIES: Pride

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			Action	DRS
	240926_LL456A1_Slot2-22_1_3139.d.zip	Other
	240926_LL456A2_Slot2-23_1_3140.d.zip	Other
	240926_LL456A3_Slot2-24_1_3141.d.zip	Other
	240926_LL456B1_Slot2-25_1_3143.d.zip	Other
	240926_LL456B2_Slot2-26_1_3144.d.zip	Other

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