Issue 4, 2016

‘Traceless’ tracing of proteins – high-affinity trans-splicing directed by a minimal interaction pair

Abstract

Protein trans-splicing mediated by split inteins is a powerful technique for site-specific protein modification. Despite recent developments there is still an urgent need for ultra-small high-affinity intein tags for in vitro and in vivo approaches. To date, only very few in-cell applications of protein trans-splicing have been reported, all limited to C-terminal protein modifications. Here, we developed a strategy for covalent N-terminal intein-mediated protein labeling at (sub) nanomolar probe concentrations. Combined with a minimal synthetic lock-and-key element, the affinity between the intein fragments was increased more than 50-fold to 10 nM. Site-specific and efficient ‘traceless’ protein modification by high-affinity trans-splicing is demonstrated at nanomolar concentrations in living mammalian cells.

Graphical abstract: ‘Traceless’ tracing of proteins – high-affinity trans-splicing directed by a minimal interaction pair

Supplementary files

Article information

Article type
Edge Article
Submitted
09 Aug 2015
Accepted
18 Dec 2015
First published
21 Dec 2015
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2016,7, 2646-2652

Author version available

‘Traceless’ tracing of proteins – high-affinity trans-splicing directed by a minimal interaction pair

M. Braner, A. Kollmannsperger, R. Wieneke and R. Tampé, Chem. Sci., 2016, 7, 2646 DOI: 10.1039/C5SC02936H

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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