Nanopore-Based Protein Identification

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.

Automated summary

This study demonstrates a method for protein detection and classification using a nanometer-scale pore formed by the protein aerolysin, achieving detection at the single-molecule level. Different water-soluble proteins treated with the same protease produce different polypeptide fragments, which can be distinguished based on their impact on the nanopore's ionic current, consistent with predicted fragments.