Reaction-based fluorogenic probes for detecting protein cysteine oxidation in living cells

Renan B. Ferreira, Ling Fu, Youngeun Jung, Jing Yang and Kate S. Carroll ()
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Renan B. Ferreira: UF Scripps Biomedical Research
Ling Fu: Beijing Institute of Lifeomics
Youngeun Jung: UF Scripps Biomedical Research
Jing Yang: Beijing Institute of Lifeomics
Kate S. Carroll: UF Scripps Biomedical Research

Nature Communications, 2022, vol. 13, issue 1, 1-12

Abstract: Abstract ‘Turn-on’ fluorescence probes for detecting H2O2 in cells are established, but equivalent tools to monitor the products of its reaction with protein cysteines have not been reported. Here we describe fluorogenic probes for detecting sulfenic acid, a redox modification inextricably linked to H2O2 signaling and oxidative stress. The reagents exhibit excellent cell permeability, rapid reactivity, and high selectivity with minimal cytotoxicity. We develop a high-throughput assay for measuring S-sulfenation in cells and use it to screen a curated kinase inhibitor library. We reveal a positive association between S-sulfenation and inhibition of TK, AGC, and CMGC kinase group members including GSK3, a promising target for neurological disorders. Proteomic mapping of GSK3 inhibitor-treated cells shows that S-sulfenation sites localize to the regulatory cysteines of antioxidant enzymes. Our studies highlight the ability of kinase inhibitors to modulate the cysteine sulfenome and should find broad application in the rapidly growing field of redox medicine.

Date: 2022
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DOI: 10.1038/s41467-022-33124-z

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