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Genetic encoding of photocaged cysteine allows photoactivation of TEV protease in live mammalian cells.

ABSTRACT: We demonstrate the evolution of the PylRS/tRNA(CUA) pair for genetically encoding photocaged cysteine. By characterizing the incorporation in Escherichia coli and mammalian cells, and the photodeprotection process in vitro and in mammalian cells, we establish conditions for rapid efficient photodeprotection to reveal native proteins in live cells. We demonstrate the utility of this approach by rapidly activating TEV protease following illumination of single cells.

SUBMITTER: Nguyen DP

PROVIDER: S-EPMC4333589 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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Genetic encoding of photocaged cysteine allows photoactivation of TEV protease in live mammalian cells.

Nguyen Duy P DP Mahesh Mohan M Elsässer Simon J SJ Hancock Susan M SM Uttamapinant Chayasith C Chin Jason W JW

Journal of the American Chemical Society 20140130 6

We demonstrate the evolution of the PylRS/tRNA(CUA) pair for genetically encoding photocaged cysteine. By characterizing the incorporation in Escherichia coli and mammalian cells, and the photodeprotection process in vitro and in mammalian cells, we establish conditions for rapid efficient photodeprotection to reveal native proteins in live cells. We demonstrate the utility of this approach by rapidly activating TEV protease following illumination of single cells. ...[more]

PMID: 24479649

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