Exploitation of single-molecule sensing by using a series of mutant aerolysin nanopores

Aerolysin, which possesses a long and narrow pore, has been widely applied to single-molecule sensing, as it could provide a nano-sized three dimensional confined space to accommodate individual biomolecules. There are abundant residues in the lumen of aerolysin that can be modulated by incorporation of different chemical group. Owing to the effect of volume exclusion and noncovalent interaction between pore and analyte on the current blockage, mutant aerolysin showed superior sensing performance. Beyond DNA detection, engineered nanopores has realized direct discrimination of adjacent phosphorylation sites in individual tau fragments and detection of Ang peptides to reveals the renin-angiotensin system crosstalk. Besides, the integration of unnatural amino acid (UAA) into aerolysin nanopore, containing aromatic groups, has the ability to distinguish peptides with hydrophobic amino acids. These well-designed aerolysin nanopores offer additional opportunities for detection of disease markers, resolving complex biological processes and even emerge as promising technology to develop single-molecule protein sequencing.