title: Free-energy calculations reveal the subtle differences in the interactions of DNA bases with α-hemolysin.
abstract: Next generation DNA sequencing methods that utilize protein nanopores have the potential to revolutionize this area of biotechnology. While the technique is underpinned by simple physics, the wild-type protein pores do not have all of the desired properties for efficient and accurate DNA sequencing. Much of the research efforts have focused on protein nanopores, such as α-hemolysin from Staphylococcus aureus. However, the speed of DNA translocation has historically been an issue, hampered in part by incomplete knowledge of the energetics of translocation. Here we have utilized atomistic molecular dynamics simulations of nucleotide fragments in order to calculate the potential of mean force (PMF) through α-hemolysin. Our results reveal specific regions within the pore that play a key role in the interaction with DNA. In particular, charged residues such as D127 and K131 provide stabilizing interactions with the anionic DNA and therefore are likely to reduce the speed of translocation. These regions provide rational targets for pore optimization. Furthermore, we show that the energetic contributions to the protein-DNA interactions are a complex combination of electrostatics and short-range interactions, often mediated by water molecules.
authors.affiliation: Chemistry, Faculty of Natural and Environmental Sciences, University of Southampton , Southampton SO17 1BJ, United Kingdom.
authors.forename: Richard M A
authors.initials: RM
authors.lastname: Manara
authors.affiliation: Chemistry, Faculty of Natural and Environmental Sciences, University of Southampton , Southampton SO17 1BJ, United Kingdom.
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Guy
authors.affiliation: Oxford Nanopore Technologies Ltd., Edmund Cartwright House, 4 Robert Robinson Avenue, Oxford Science Park, Oxford OX4 4GA, United Kingdom.
authors.forename: E Jayne
authors.initials: EJ
authors.lastname: Wallace
authors.affiliation: Chemistry, Faculty of Natural and Environmental Sciences, University of Southampton , Southampton SO17 1BJ, United Kingdom.
authors.forename: Syma
authors.initials: S
authors.lastname: Khalid
mesh_terms: D020029:Base Pairing; D004247:DNA / Q000737:chemistry*; D006460:Hemolysin Proteins / Q000737:chemistry*; D056004:Molecular Dynamics Simulation; D013211:Staphylococcus aureus / Q000737:chemistry; D055672:Static Electricity; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
chemical_list: D006460:Hemolysin Proteins; D014867:Water; D004247:DNA