Mapping the sensing spots of aerolysin for single oligonucleotides analysis

Chan Cao, Meng-Yin Li, Nuria Cirauqui, Ya-Qian Wang, Matteo Dal Peraro (), He Tian and Yi-Tao Long ()
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Chan Cao: East China University of Science and Technology
Meng-Yin Li: East China University of Science and Technology
Nuria Cirauqui: School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL)
Ya-Qian Wang: East China University of Science and Technology
Matteo Dal Peraro: School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL)
He Tian: East China University of Science and Technology
Yi-Tao Long: East China University of Science and Technology

Nature Communications, 2018, vol. 9, issue 1, 1-9

Abstract: Abstract Nanopore sensing is a powerful single-molecule method for DNA and protein sequencing. Recent studies have demonstrated that aerolysin exhibits a high sensitivity for single-molecule detection. However, the lack of the atomic resolution structure of aerolysin pore has hindered the understanding of its sensing capabilities. Herein, we integrate nanopore experimental results and molecular simulations based on a recent pore structural model to precisely map the sensing spots of this toxin for ssDNA translocation. Rationally probing ssDNA length and composition upon pore translocation provides new important insights for molecular determinants of the aerolysin nanopore. Computational and experimental results reveal two critical sensing spots (R220, K238) generating two constriction points along the pore lumen. Taking advantage of the sensing spots, all four nucleobases, cytosine methylation and oxidation of guanine can be clearly identified in a mixture sample. The results provide evidence for the potential of aerolysin as a nanosensor for DNA sequencing.

Date: 2018
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DOI: 10.1038/s41467-018-05108-5

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