Nanopore-Based Protein Identification

作者：Mazdak Afshar Bakshloo, John J. Kasianowicz, Manuela Pastoriza‐Gallego, Jérôme Mathé, Régis Daniel, Fabien Piguet, Abdelghani Oukhaled · 发表于：Journal of the American Chemical Society · 年份：2022 · DOI：10.1021/jacs.1c11758 · 被引用次数：150 · 研究领域：Nanopore and Nanochannel Transport Studies、Ion-surface interactions and analysis、Microfluidic and Capillary Electrophoresis Applications

The implementation of a reliable, rapid, inexpensive, and simple method for whole-proteome identification would greatly benefit cell biology research and clinical medicine. Proteins are currently identified by cleaving them with proteases, detecting the polypeptide fragments with mass spectrometry, and mapping the latter to sequences in genomic/proteomic databases. Here, we demonstrate that the polypeptide fragments can instead be detected and classified at the single-molecule limit using a nanometer-scale pore formed by the protein aerolysin. Specifically, three different water-soluble proteins treated with the same protease, trypsin, produce different polypeptide fragments defined by the degree by which the latter reduce the nanopore's ionic current. The fragments identified with the aerolysin nanopore are consistent with the predicted fragments that trypsin could produce.