title: Does the lipid environment impact the open-state conductance of an engineered β-barrel protein nanopore?
abstract: Using rational membrane protein design, we were recently able to obtain a β-barrel protein nanopore that was robust under an unusually broad range of experimental circumstances. This protein nanopore was based upon the native scaffold of the bacterial ferric hydroxamate uptake component A (FhuA) of Escherichia coli. In this work, we expanded the examinations of the open-state current of this engineered protein nanopore, also called FhuA ΔC/Δ4L, employing an array of lipid bilayer systems that contained charged and uncharged as well as conical and cylindrical lipids. Remarkably, systematical single-channel analysis of FhuA ΔC/Δ4L indicated that most of its biophysical features, such as the unitary conductance and the stability of the open-state current, were not altered under the conditions tested in this work. However, electrical recordings at high transmembrane potentials revealed that the presence of conical phospholipids within the bilayer catalyzes the first, stepwise current transition of the FhuA ΔC/Δ4L protein nanopore to a lower-conductance open state. This study reinforces the stability of the open-state current of the engineered FhuA ΔC/Δ4L protein nanopore under various experimental conditions, paving the way for further critical developments in biosensing and molecular biomedical diagnosis.
authors.affiliation: Department of Physics, Syracuse University, Syracuse, NY, USA.
authors.forename: Noriko
authors.initials: N
authors.lastname: Tomita
authors.forename: Mohammad M
authors.initials: MM
authors.lastname: Mohammad
authors.forename: David J
authors.initials: DJ
authors.lastname: Niedzwiecki
authors.forename: Makoto
authors.initials: M
authors.lastname: Ohta
authors.forename: Liviu
authors.initials: L
authors.lastname: Movileanu
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D001703:Biophysics / Q000379:methods; D015374:Biosensing Techniques; D004553:Electric Conductivity; D004594:Electrophysiology; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D008564:Membrane Potentials; D008968:Molecular Conformation; D053758:Nanoparticles / Q000737:chemistry; D058608:Nanopores; D036103:Nanotechnology / Q000379:methods; D015202:Protein Engineering / Q000379:methods*; D017433:Protein Structure, Secondary
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; C050900:FhuA protein, E coli; D008051:Lipid Bilayers; D008055:Lipids