Bioorthogonal photocatalytic proximity labeling in primary living samples

Ziqi Liu, Fuhu Guo, Yufan Zhu, Shengnan Qin, Yuchen Hou, Haotian Guo, Feng Lin, Peng R. Chen () and Xinyuan Fan ()
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Ziqi Liu: Peking University
Fuhu Guo: Peking University
Yufan Zhu: Peking University
Shengnan Qin: Peking University
Yuchen Hou: Peking University
Haotian Guo: Peking University
Feng Lin: Peking University
Peng R. Chen: Peking University
Xinyuan Fan: Peking University

Nature Communications, 2024, vol. 15, issue 1, 1-18

Abstract: Abstract In situ profiling of subcellular proteomics in primary living systems, such as native tissues or clinic samples, is crucial for understanding life processes and diseases, yet challenging due to methodological obstacles. Here we report CAT-S, a bioorthogonal photocatalytic chemistry-enabled proximity labeling method, that expands proximity labeling to a wide range of primary living samples for in situ profiling of mitochondrial proteomes. Powered by our thioQM labeling warhead development and targeted bioorthogonal photocatalytic chemistry, CAT-S enables the labeling of mitochondrial proteins in living cells with high efficiency and specificity. We apply CAT-S to diverse cell cultures, dissociated mouse tissues as well as primary T cells from human blood, portraying the native-state mitochondrial proteomic characteristics, and unveiled hidden mitochondrial proteins (PTPN1, SLC35A4 uORF, and TRABD). Furthermore, CAT-S allows quantification of proteomic perturbations on dysfunctional tissues, exampled by diabetic mouse kidneys, revealing the alterations of lipid metabolism that may drive disease progression. Given the advantages of non-genetic operation, generality, and spatiotemporal resolution, CAT-S may open exciting avenues for subcellular proteomic investigations of primary samples that are otherwise inaccessible.

Date: 2024
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DOI: 10.1038/s41467-024-46985-3

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