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Issue 4, 2016
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‘Traceless’ tracing of proteins – high-affinity trans-splicing directed by a minimal interaction pair

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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

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Publication details

The article was received on 09 Aug 2015, accepted on 18 Dec 2015 and first published on 21 Dec 2015


Article type: Edge Article
DOI: 10.1039/C5SC02936H
Citation: Chem. Sci., 2016,7, 2646-2652
  • Open access: Creative Commons BY license
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    ‘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

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