Bioorthogonal Photocatalytic Decaging-Enabled Mitochondrial Proteomics.
Zongyu HuangZiqi LiuXiao XieRuxin ZengZujie ChenLinghao KongXinyuan FanPeng R ChenPublished in: Journal of the American Chemical Society (2021)
Spatiotemporally resolved dissection of subcellular proteome is crucial to our understanding of cellular functions in health and disease. We herein report a bioorthogonal and photocatalytic decaging-enabled proximity labeling strategy (CAT-Prox) for spatiotemporally resolved mitochondrial proteome profiling in living cells. Our systematic survey of the photocatalysts has led to the identification of Ir(ppy)2bpy as a bioorthogonal and mitochondria-targeting catalyst that allowed photocontrolled, rapid rescue of azidobenzyl-caged quinone methide as a highly reactive Michael acceptor for proximity-based protein labeling in mitochondria of live cells. Upon careful validation through in vitro labeling, mitochondria-targeting specificity, in situ catalytic activity as well as protein tagging, we applied CAT-Prox for mitochondria proteome profiling in living Hela cells as well as hard-to-transfect macrophage RAW264.7 cells with approximately 70% mitochondria specificity observed from up to 300 proteins enriched. Finally, CAT-Prox was further applied to the dynamic dissection of mitochondria proteome of macrophage cells upon lipopolysaccharide stimulation. By integrating photocatalytic decaging chemistry with proximity-based protein labeling, CAT-Prox offers a general, catalytic, and nongenetic alternative to the enzyme-based proximity labeling strategies for diverse live cell settings.
Keyphrases
- induced apoptosis
- cell cycle arrest
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- oxidative stress
- living cells
- reactive oxygen species
- endoplasmic reticulum stress
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- reduced graphene oxide
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- gold nanoparticles
- toll like receptor
- climate change
- amino acid
- pi k akt
- cross sectional
- cell proliferation
- metal organic framework
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- carbon dioxide