Protein labeling in living cells - A combination of enzyme mediated peptide labeling and inverse electron demand Diels-Alder cycloaddition

In vivo protein labeling is to date the most common approach to study a protein’s function in real time and in its native environment. We develop a protein labeling strategy that will make significant impact by providing a minimally disruptive technology (minimal tag size) for labeling proteins selectively in live cells. The key elements of the new labeling technology are enzyme mediated peptide labeling and the highly bioorthogonal inverse electron demand Diels-Alder reaction.

This strategy enables the introduction of small reporter molecules that match the specific needs for different biophysical imaging techniques in live cells, like superresolution microscopy or single molecule imaging. We further expand the new labeling approach to a protein mislocalization approach which will enable a fast and acute regulation of protein activity in live cells to study cellular processes.

Dr. Richard Wombacher
Ruprechts-Karls Universität Heidelberg

Tel.: +49 6221 54 4879
Fax: +49 6221 54 6430

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Publications within the SPP 1623 project

A. Wieczorek, P. Werther, J. Euchner, R. Wombacher
2017, 8, 1506-1510
Green- to far-red-emitting fluorogenic tetrazine probes - synthetic access and no-wash protein imaging inside living cells.
Link to the article

S. Hauke, A. von Appen, T. Quidwar, J. Ries, R. Wombacher
2017, 8(1), 559-566
Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells.
Link to the article

M. Best, I. Porth, S. Hauke, F. Braun, D.P. Herten, R. Wombacher
Org. Biomol. Chem
2016, 14(24), 5606-5611
Protein-specific localization of a rhodamine-based calcium-sensor in living cells.
Link to the article

M. Best, A. Degen, T.T. Schmidt, M. Baalmann, R. Wombacher
2015, 16, 1158-1162.
Two-Step Protein Labeling by Using Lipoic Acid Ligase with Norbornene Substrates and Subsequent Inverse-Electron Demand Diels-Alder Reaction.
Link to the article

K.M. Schelkle, T. Griesbaum, D. Ollech, S. Becht, T. Buckup, M. Hamburger, R. Wombacher
Angew. Chem. Int. Ed. 2015, 54, 2825-2829
Light-Induced Protein Dimerization by One- and Two-Photon Activation of Gibberellic Acid.

S. Hauke, M. Best, T.T. Schmidt, M. Baalmann. A. Krause, R. Wombacher
, 2014, 25(9), 1632-1637.
Two-Step Protein labeling Utilizing Lipoic Acid Ligase and Sonogashira Cross-Coupling
Link to the article

A. Wieczorek, T. Buckup, R. Wombacher
2014, 12 (24), 4177-4185
Rigid tetrazine fluorophore conjugates with fluorogenic properties in inverse electron demand Diels-Alder reaction
Link to the article