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Label-Free and Real-Time Detection of Protein Ubiquitination with a Biological Nanopore.

ABSTRACT: The covalent addition of ubiquitin to target proteins is a key post-translational modification that is linked to a myriad of biological processes. Here, we report a fast, single-molecule, and label-free method to probe the ubiquitination of proteins employing an engineered Cytolysin A (ClyA) nanopore. We show that ionic currents can be used to recognize mono- and polyubiquitinated forms of native proteins under physiological conditions. Using defined conjugates, we also show that isomeric monoubiquitinated proteins can be discriminated. The nanopore approach allows following the ubiquitination reaction in real time, which will accelerate the understanding of fundamental mechanisms linked to protein ubiquitination.

SUBMITTER: Wloka C

PROVIDER: S-EPMC5444049 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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Label-Free and Real-Time Detection of Protein Ubiquitination with a Biological Nanopore.

Wloka Carsten C Van Meervelt Veerle V van Gelder Dewi D Danda Natasha N Jager Nienke N Williams Chris P CP Maglia Giovanni G

ACS nano 20170329 5

The covalent addition of ubiquitin to target proteins is a key post-translational modification that is linked to a myriad of biological processes. Here, we report a fast, single-molecule, and label-free method to probe the ubiquitination of proteins employing an engineered Cytolysin A (ClyA) nanopore. We show that ionic currents can be used to recognize mono- and polyubiquitinated forms of native proteins under physiological conditions. Using defined conjugates, we also show that isomeric monoub ...[more]

PMID: 28353339

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