Alpha‐Helical Fragaceatoxin C Nanopore Engineered for Double‐Stranded and Single‐Stranded Nucleic Acid Analysis |
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| Authors: | Dr Carsten Wloka Natalie Lisa Mutter Dr Misha Soskine Prof Dr Giovanni Maglia |
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| Affiliation: | Chemical Biology I, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Groningen, The Netherlands |
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| Abstract: | Nanopores are used in single‐molecule DNA analysis and sequencing. Herein, we show that Fragaceatoxin C (FraC), an α‐helical pore‐forming toxin from an actinoporin protein family, can be reconstituted in sphingomyelin‐free standard planar lipid bilayers. We engineered FraC for DNA analysis and show that the funnel‐shaped geometry allows tight wrapping around single‐stranded DNA (ssDNA), resolving between homopolymeric C, T, and A polynucleotide stretches. Remarkably, despite the 1.2 nm internal constriction of FraC, double‐stranded DNA (dsDNA) can translocate through the nanopore at high applied potentials, presumably through the deformation of the α‐helical transmembrane region of the pore. Therefore, FraC nanopores might be used in DNA sequencing and dsDNA analysis. |
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| Keywords: | DNA analysis DNA sequencing fragaceatoxins nanopores single-molecule studies |
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