Small-molecule control of protein function through staudinger reduction

ABSTRACT Using small molecules to control the function of proteins in live cells with complete specificity is highly desirable, but challenging. Here we report a small-molecule switch that

can be used to control protein activity. The approach uses a phosphine-mediated Staudinger reduction to activate protein function. Genetic encoding of an _ortho_-azidobenzyloxycarbonyl amino

acid using a pyrrolysyl transfer RNA synthetase/tRNACUA pair in mammalian cells enables the site-specific introduction of a small-molecule-removable protecting group into the protein of

interest. Strategic placement of this group renders the protein inactive until deprotection through a bioorthogonal Staudinger reduction delivers the active wild-type protein. This developed

methodology was applied to the conditional control of several cellular processes, including bioluminescence (luciferase), fluorescence (enhanced green fluorescent protein), protein

translocation (nuclear localization sequence), DNA recombination (Cre) and gene editing (Cas9). Access through your institution Buy or subscribe This is a preview of subscription content,

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DECAGING IN LIVING CELLS Article 28 February 2024 METAL-RESPONSIVE REGULATION OF ENZYME CATALYSIS USING GENETICALLY ENCODED CHEMICAL SWITCHES Article Open access 06 April 2022 A SPLIT

RIBOZYME THAT LINKS DETECTION OF A NATIVE RNA TO ORTHOGONAL PROTEIN OUTPUTS Article Open access 01 February 2023 ACCESSION CODES PRIMARY ACCESSIONS PROTEIN DATA BANK * 1CRX * 2D1S * 4EUL

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(2015). CAS  PubMed  PubMed Central  Google Scholar  Download references ACKNOWLEDGEMENTS This research was supported in part by the National Institutes of Health (1R01GM112728), the

National Science Foundation (MCB-1330746) and the Charles E. Kaufman Foundation of The Pittsburgh Foundation. K.M. is grateful for a Japan Society for the Promotion of Science Postdoctoral

Fellowship for Research Abroad. We thank the Chin lab (Medical Research Council) for plasmids encoding the _PylRS_ and _PylT_ genes, the Asokan lab (University of North Carolina) for the

pgRNA and pIRG plasmids and the Hughes lab (Montana State University) for the Cre Stoplight plasmid. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Chemistry, University of

Pittsburgh, Pittsburgh, 15260, Pennsylvania, USA Ji Luo, Qingyang Liu, Kunihiko Morihiro & Alexander Deiters Authors * Ji Luo View author publications You can also search for this author

inPubMed Google Scholar * Qingyang Liu View author publications You can also search for this author inPubMed Google Scholar * Kunihiko Morihiro View author publications You can also search

for this author inPubMed Google Scholar * Alexander Deiters View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS A.D. and J.L. conceived and

designed the experiments. J.L, Q.L. and K.M. performed the experiments and analysed the data. A.D. and J.L. co-wrote the paper. CORRESPONDING AUTHOR Correspondence to Alexander Deiters.

ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary information (PDF 2316 kb)

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CITE THIS ARTICLE Luo, J., Liu, Q., Morihiro, K. _et al._ Small-molecule control of protein function through Staudinger reduction. _Nature Chem_ 8, 1027–1034 (2016).

https://doi.org/10.1038/nchem.2573 Download citation * Received: 11 November 2015 * Accepted: 13 June 2016 * Published: 25 July 2016 * Issue Date: November 2016 * DOI:

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