title: Single-molecule transport across an individual biomimetic nuclear pore complex.
abstract: Nuclear pore complexes regulate the selective exchange of RNA and proteins across the nuclear envelope in eukaryotic cells. Biomimetic strategies offer new opportunities to investigate this remarkable transport phenomenon. Here, we show selective transport of proteins across individual biomimetic nuclear pore complexes at the single-molecule level. Each biomimetic complex is constructed by covalently tethering either Nup98 or Nup153 (phenylalanine-glycine (FG) nucleoporins) to a solid-state nanopore. Individual translocation events are monitored using ionic current measurements with sub-millisecond temporal resolution. Transport receptors (Impβ) proceed with a dwell time of ∼2.5 ms for both Nup98- and Nup153-coated pores, whereas the passage of non-specific proteins is strongly inhibited with different degrees of selectivity. For pores up to ∼25 nm in diameter, Nups form a dense and low-conducting barrier, whereas they adopt a more open structure in larger pores. Our biomimetic nuclear pore complex provides a quantitative platform for studying nucleocytoplasmic transport phenomena at the single-molecule level in vitro.
authors.affiliation: Kavli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands.
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Kowalczyk
authors.forename: Larisa
authors.initials: L
authors.lastname: Kapinos
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Blosser
authors.forename: Tomás
authors.initials: T
authors.lastname: Magalhães
authors.forename: Pauline
authors.initials: P
authors.lastname: van Nies
authors.forename: Roderick Y H
authors.initials: RY
authors.lastname: Lim
authors.forename: Cees
authors.initials: C
authors.lastname: Dekker
mesh_terms: D040761:Biomimetic Materials / Q000737:chemistry* / Q000378:metabolism; D004553:Electric Conductivity; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D022022:Nuclear Pore / Q000737:chemistry* / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry / Q000378:metabolism; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D028861:Nuclear Pore Complex Proteins; D011506:Proteins; D011994:Recombinant Proteins