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Genetically encoding thyronine for fluorescent detection of peroxynitrite.

ABSTRACT: Peroxynitrite is a highly reactive oxidant effecting cell signaling and cell death. Here we report a fluorescent protein probe to selectively detect peroxynitrite. A novel unnatural amino acid, thyronine (Thy), was genetically encoded in E. coli and mammalian cells by evolving an orthogonal tRNA^Pyl/ThyRS pair. Incorporation of Thy into the chromophore of sfGFP or cpsGFP afforded a virtually non-fluorescent reporter. Upon treatment with peroxynitrite, Thy was converted into tyrosine via O-dearylation, regenerating GFP fluorescence in a time- and concentration-dependent manner. Genetically encoded thyronine may also be valuable for other redox applications.

SUBMITTER: Li S

PROVIDER: S-EPMC7461609 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Genetically encoding thyronine for fluorescent detection of peroxynitrite.

Li Shanshan S Yang Bing B Kobayashi Tomonori T Yu Bingchen B Liu Jun J Wang Lei L

Bioorganic & medicinal chemistry 20200729 18

Peroxynitrite is a highly reactive oxidant effecting cell signaling and cell death. Here we report a fluorescent protein probe to selectively detect peroxynitrite. A novel unnatural amino acid, thyronine (Thy), was genetically encoded in E. coli and mammalian cells by evolving an orthogonal tRNA<sup>Pyl</sup>/ThyRS pair. Incorporation of Thy into the chromophore of sfGFP or cpsGFP afforded a virtually non-fluorescent reporter. Upon treatment with peroxynitrite, Thy was converted into tyrosine vi ...[more]

PMID: 32828428

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