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

P. Werther, J.S. Möhler, R. Wombacher
Chem. Eur. J. 2017, 72, 18216-18224
A Bifunctional Fluorogenic Rhodamine Probe for Proximity-Induced Bioorthogonal Chemistry.
Link to the article

A. Wieczorek, P. Werther, J. Euchner, R. Wombacher
Chem.Sci. 2017, 8, 1506-1510
Green- to far-red-emitting fluorogenic tetrazine probes - synthetic access and no-wash protein imaging inside living cells.
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S. Hauke, A. von Appen, T. Quidwar, J. Ries, R. Wombacher
Chem.Sci. 2017, 8(1), 559-566
Specific protein labeling with caged fluorophores for dual-color imaging and super-resolution microscopy in living cells.
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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