Label-Free and Real-Time Detection of Protein Ubiquitination with a Biological Nanopore.
Label-Free and Real-Time Detection of Protein Ubiquitination with a Biological Nanopore.
复制标题
生物纳米孔蛋白泛素化的无标记实时检测。
DOI：
10.1021/acsnano.6b07760
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发表时间：
2017-05-23
期刊：
影响因子：
17.1
通讯作者：
Maglia G
中科院分区：
文献类型：
--
作者：
Wloka C;Van Meervelt V;van Gelder D;Danda N;Jager N;Williams CP;Maglia G
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.
