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Photocatalytic Golgi Proteomics Reveals Palmitoylation-Regulated Golgiphagy

ABSTRACT: The Golgi apparatus (GA) orchestrates protein modification, trafficking, and secretion through highly dynamic remodel-ing, yet its proteomic complexity remains difficult to resolve in living systems. Here, we report CAT-Golgi, a genetically independent and light-controlled photocatalytic proximity labeling strategy for in situ spatiotemporal mapping of the Golgi proteome. Combining a cysteine-conjugated eosin photocatalyst (GolgiCat) with an aniline probe, CAT-Golgi enables rapid and precise protein labeling within minutes under mild green light, requiring no genetic manipulation and operat-ing efficiently in hard-to-transfect and primary cells. Leveraging our extensive efforts in organelle-targeted photocatalytic systems, we extended this chemistry to the highly dynamic and reversible Golgi apparatus. CAT-Golgi achieved quantita-tive and comparative proteomics in HeLa, K562, Jurkat and primary HEKa cells, revealing both conserved and cell-type-specific profiles. Under Brefeldin A-induced Golgiphagy, CAT-Golgi captured large-scale proteome remodeling and identi-fied palmitoyl-protein thioesterase 1 (PPT1) as a central regulator. PPT1 downregulation enhanced ULK1 and TRPML1 palmitoylation, disrupted redox balance, and activated Golgiphagy. CAT-Golgi provides a broadly applicable chemical platform for decoding organelle dynamics, offering both conceptual and technical foundations for extending photocata-lytic proteomics to other transient organelles and illuminating molecular mechanisms of organelle plasticity and disease progression.

ORGANISM(S): Homo Sapiens

SUBMITTER: Xinyuan Fan

PROVIDER: PXD075800 | iProX | Wed Mar 18 00:00:00 GMT 2026

REPOSITORIES: iProX

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