Single-molecule sensing of peptides and nucleic acids by engineered aerolysin nanopores

Author:

Cao ChanORCID,Cirauqui NuriaORCID,Marcaida Maria Jose,Buglakova Elena,Duperrex Alice,Radenovic AleksandraORCID,Dal Peraro Matteo

Abstract

Abstract Nanopore sensing is a powerful single-molecule approach for the detection of biomolecules. Recent studies have demonstrated that aerolysin is a promising candidate to improve the accuracy of DNA sequencing and to develop novel single-molecule proteomic strategies. However, the structure–function relationship between the aerolysin nanopore and its molecular sensing properties remains insufficiently explored. Herein, a set of mutated pores were rationally designed and evaluated in silico by molecular simulations and in vitro by single-channel recording and molecular translocation experiments to study the pore structural variation, ion selectivity, ionic conductance and capabilities for sensing several biomolecules. Our results show that the ion selectivity and sensing ability of aerolysin are mostly controlled by electrostatics and the narrow diameter of the double β-barrel cap. By engineering single-site mutants, a more accurate molecular detection of nucleic acids and peptides has been achieved. These findings open avenues for developing aerolysin nanopores into powerful sensing devices.

Funder

École Polytechnique Fédérale de Lausanne

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung

Novartis Stiftung für Medizinisch-Biologische Forschung

Publisher

Springer Science and Business Media LLC

Subject

General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry

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