title: Controlled translocation of individual DNA molecules through protein nanopores with engineered molecular brakes.
abstract: Protein nanopores may provide a cheap and fast technology to sequence individual DNA molecules. However, the electrophoretic translocation of ssDNA molecules through protein nanopores has been too rapid for base identification. Here, we show that the translocation of DNA molecules through the α-hemolysin protein nanopore can be slowed controllably by introducing positive charges into the lumen of the pore by site directed mutagenesis. Although the residual ionic current during DNA translocation is insufficient for direct base identification, we propose that the engineered pores might be used to slow down DNA in hybrid systems, for example, in combination with solid-state nanopores.
authors.affiliation: Department of Chemistry, University of Oxford, Oxford, United Kingdom.
authors.forename: Marcela
authors.initials: M
authors.lastname: Rincon-Restrepo
authors.forename: Ellina
authors.initials: E
authors.lastname: Mikhailova
authors.forename: Hagan
authors.initials: H
authors.lastname: Bayley
authors.forename: Giovanni
authors.initials: G
authors.lastname: Maglia
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*; D008567:Membranes, Artificial; D009038:Motion; D009154:Mutation; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure*; D016062:Porosity; D011485:Protein Binding; D013329:Structure-Activity Relationship
chemical_list: D004268:DNA-Binding Proteins; D008567:Membranes, Artificial; D004247:DNA