NPC mimics: probing the mechanism of nucleocytoplasmic transport

Methods Cell Biol. 2014:122:379-93. doi: 10.1016/B978-0-12-417160-2.00017-5.

Abstract

In vitro mimics of cellular machines have been recently engineered and utilized to investigate processes within cells. These devices can provide novel insights into biological mechanisms and have the potential to improve biotechnological processes such as separation. In particular, several devices have been designed to mimic translocation through nuclear pore complexes (NPCs). We describe here the fabrication of a biomimetic NPC using nanoporous filters lined with FG-repeats of proteins that create a selectivity barrier. We show the utility of this nanoselective filter as a testbed for the investigation of nucleocytoplasmic transport and demonstrate that this device closely reproduces key features of trafficking through the NPC.

Keywords: Flux measurements; NPC; NPC mimics; Nanopores; Nucleocytoplasmic transport; Permeability; Transport kinetics.

MeSH terms

  • Active Transport, Cell Nucleus / physiology*
  • Bacterial Proteins
  • Biomimetic Materials / metabolism*
  • Glutathione Transferase / chemistry
  • Gold / chemistry
  • Lipid Bilayers / chemistry
  • Luminescent Proteins
  • Microscopy, Confocal / methods
  • Models, Biological
  • Muramidase / metabolism
  • Nanopores
  • Nuclear Pore / metabolism*
  • Nuclear Pore Complex Proteins / genetics
  • Nuclear Pore Complex Proteins / metabolism
  • Nucleocytoplasmic Transport Proteins / metabolism
  • Perforin / chemistry
  • Polycarboxylate Cement / chemistry
  • Pregnancy Proteins / metabolism
  • Protein Transport / physiology*
  • Saccharomyces cerevisiae
  • Staining and Labeling
  • Thrombin / metabolism
  • beta Karyopherins
  • ran GTP-Binding Protein / metabolism

Substances

  • Bacterial Proteins
  • Lipid Bilayers
  • Luminescent Proteins
  • NUTF2 protein, human
  • Nuclear Pore Complex Proteins
  • Nucleocytoplasmic Transport Proteins
  • Polycarboxylate Cement
  • Pregnancy Proteins
  • beta Karyopherins
  • yellow fluorescent protein, Bacteria
  • Perforin
  • polycarbonate
  • Gold
  • Glutathione Transferase
  • Muramidase
  • Thrombin
  • ran GTP-Binding Protein