Multi-scale photocatalytic proximity labeling reveals cell surface neighbors on and between cells

ABSTRACT: Proximity labeling proteomics (PLP) strategies are powerful approaches to yield snapshots of protein neighborhoods. Here, we describe a multi-scale PLP method with adjustable resolution using a commercially available photosensitizer, Eosin Y, which upon visible light illumination, activates different photo-probes with a range of labeling radii. We applied this platform to profile neighborhoods of the oncogenic epidermal growth factor receptor (EGFR) and orthogonally validated >20 neighbors using immuno-assays and AlphaFold-Multimer prediction. We further profiled the protein neighborhoods of cell-cell synapses induced by bi-specific T-cell engagers (BiTEs) and chimeric antigen receptor (CAR)T cells. This integrated multi-scale PLP platform maps local and distal protein networks on and between cell surfaces, which will aid in the systematic construction of the cell surface interactome and reveal new immunotherapeutic opportunities.

INSTRUMENT(S): timsTOF Pro, timsTOF Pro 2

ORGANISM(S): Acyrthosiphon Pisum (pea Aphid) Homo Sapiens (human)

SUBMITTER: Zhi Lin  

LAB HEAD: James A. Wells

PROVIDER: PXD051348 | Pride | 2025-02-17

REPOSITORIES: Pride