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

Carsten Wloka, Veerle Van Meervelt, Dewi van Gelder, Natasha Danda, Nienke Jager, Chris P Williams, Giovanni Maglia

OnderzoeksoutputAcademicpeer review

72 Citaten (Scopus)
433 Downloads (Pure)

Samenvatting

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.

Originele taal-2English
Artikelnummeracsnano.6b07760
Pagina's (van-tot)4387-4394
Aantal pagina's8
TijdschriftAcs Nano
Volume11
Nummer van het tijdschrift5
Vroegere onlinedatummrt-2017
DOI's
StatusPublished - apr-2017

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