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'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.

SUBMITTER: Braner M

PROVIDER: S-EPMC5477019 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature

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'Traceless' tracing of proteins - high-affinity trans-splicing directed by a minimal interaction pair.

Braner M M Kollmannsperger A A Wieneke R R Tampé R R

Chemical science 20151221 4

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 <i>in vitro</i> and <i>in vivo</i> 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) n ...[more]

PMID: 28660037

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