Specific labeling of proteins within living cells with single residue precision through genetically encoded click chemistry

The development of catalyst-free click chemistry, i.e. a chemical reaction that is non-toxic and proceeds readily and specifically even within living organisms, has revolutionized chemical biology. By encoding strained alkyne unnatural amino acids into the genetic repertoire of the cell, we exploited this chemistry to label proteins with single residue specificity in bacteria. Meanwhile, we extended the method to more click type reactions, creating a unique opportunity to establish these techniques for installing probes to control and visualize protein function within living species.

In this proposal, we aim to lay the foundation for a smaller and more stable substitute to fluorescent proteins in biology, medicine, and diagnostics. In particular, we aim to explore the potential of click chemistries of the Huisgen-type and inverse electron-demand Diels-Alder reaction and optimize and characterize probes in vitro and in vivo. We will use the techniques to study growth factor receptor signaling and to develop genetically encoded small molecule dye FRET sensors

Dr. Edward Lemke
European Molecular Biology Laboratory (EMBL)

Tel.: +49 6221 387 - 8526

Email Dr. Lemke

Dr. Carsten Schultz
European Molecular Biology Laboratory (EMBL)

Email Dr. Schultz

Publications within the SPP 1623 project

J.A. Wagner, D. Mercadante, I. Nikic, E.A. Lemke, F. Gräter
Chem. Eur. J. 2015 , 21(35), 12431-12435
Orthogonality of Strain-Promoted Click Reactions
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J.E. Hoffmann, T. Plass, I. Nikic, I.V. Aramburu, C. Köhler, H. Gillandt, E.A. Lemke, C. Schultz
Chemistry 2015, 21(35), 12266-12270
Highly stable trans-cyclooctene amino acids for live cell labeling
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S. Tyagi, E.A. Lemke
Curr. Opin. Struct. Biol. 2015, 32, 66-73
Single-molecule FRET and crosslinking studies in structural biology enabled by noncanonical amino acids.
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I. Nikic, T. Plass, O. Schraidt, J. Szymanski, J.A. Briggs, C. Schultz, E.A. Lemke
Angew. Chem. Int. Ed.2014, 53 (8), 2245-2249
Minimal Tags for Rapid Dual-Color Live-Cell Labeling and Super-Resolution Microscopy
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E.A. Lemke
Chembiochem. 2014, 15(12), 1691-1694
The exploding genetic code.
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A. Herner, I. Nikic, M. Kallay, E.A. Lemke, P. Kele
Org Biomol Chem 2013, 11(20), 3297-3306
A new family of bioorthogonally applicable fluorogenic labels.
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S. Milles, E.A. Lemke
Bioessays2013, 35(1), 65-74
What precision-protein-tuning and nano-resolved single molecule sciences can do for each other
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