Genetic Encoding of Photocaged Cysteine Allows Photoactivation of TEV Protease in Live Mammalian Cells

Nguyen DP, Mahesh M, Elsässer SJ, Hancock SM, Uttamapinant C, Chin JW

J. Am. Chem. Soc. 136 (6) 2240-2243 [2014-02-12; online 2014-01-30]

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.

Simon Elsässer

PubMed 24479649

DOI 10.1021/ja412191m

Crossref 10.1021/ja412191m