Electro-Osmotic Vortices Promote the Capture of Folded Proteins by PlyAB Nanopores

Gang Huang, Kherim Willems, Mart Bartelds, Pol van Dorpe, Misha Soskine, Giovanni Maglia*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)
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Abstract

Biological nanopores are emerging as powerful tools for single-molecule analysis and sequencing. Here, we engineered the two-component pleurotolysin (PlyAB) toxin to assemble into 7.2 × 10.5 nm cylindrical nanopores with a low level of electrical noise in lipid bilayers, and we addressed the nanofluidic properties of the nanopore by continuum simulations. Surprisingly, proteins such as human albumin (66.5 kDa) and human transferrin (76-81 kDa) did not enter the nanopore. We found that the precise engineering of the inner surface charge of the PlyAB induced electro-osmotic vortices that allowed the electrophoretic capture of the proteins. Once inside the nanopore, two human plasma proteins could be distinguished by the characteristics of their current blockades. This fundamental understanding of the nanofluidic properties of nanopores provides a practical method to promote the capture and analysis of folded proteins by nanopores.

Original languageEnglish
Pages (from-to)3819-3827
Number of pages9
JournalNano Letters
Volume20
Issue number5
Early online date9-Apr-2020
DOIs
Publication statusPublished - 13-May-2020

Keywords

  • biological nanopore
  • single-molecule sensing
  • electro-osmosis
  • plasma proteins
  • nanofluidics
  • SOLID-STATE NANOPORES
  • ALPHA-HEMOLYSIN
  • SINGLE
  • TRANSLOCATION
  • PEPTIDES
  • SIZE
  • MOLECULES
  • POLYMER
  • DISCRIMINATION
  • IDENTIFICATION

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