title: NPC mimics: probing the mechanism of nucleocytoplasmic transport.
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.
authors.affiliation: Department of Molecular Medicine, Beckman Research Institute of the City of Hope Comprehensive Cancer Center, Duarte, California, USA.
authors.forename: Tijana
authors.initials: T
authors.lastname: Jovanovic-Talisman
authors.affiliation: Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, USA.
authors.forename: Brian T
authors.initials: BT
authors.lastname: Chait
authors.affiliation: Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Rout
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D001426:Bacterial Proteins; D040761:Biomimetic Materials / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D008164:Luminescent Proteins; D018613:Microscopy, Confocal / Q000379:methods; D008954:Models, Biological; D009113:Muramidase / Q000378:metabolism; D058608:Nanopores; D022022:Nuclear Pore / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism; D054353:Perforin / Q000737:chemistry; D011076:Polycarboxylate Cement / Q000737:chemistry; D011257:Pregnancy Proteins / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology*; D012441:Saccharomyces cerevisiae; D013194:Staining and Labeling; D013917:Thrombin / Q000378:metabolism; D028961:beta Karyopherins; D020931:ran GTP-Binding Protein / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008051:Lipid Bilayers; D008164:Luminescent Proteins; C038965:NUTF2 protein, human; D028861:Nuclear Pore Complex Proteins; D029543:Nucleocytoplasmic Transport Proteins; D011076:Polycarboxylate Cement; D011257:Pregnancy Proteins; D028961:beta Karyopherins; C054562:yellow fluorescent protein, Bacteria; D054353:Perforin; C005506:polycarbonate; D006046:Gold; D005982:Glutathione Transferase; D009113:Muramidase; D013917:Thrombin; D020931:ran GTP-Binding Protein
keywords: Flux measurements; NPC; NPC mimics; Nanopores; Nucleocytoplasmic transport; Permeability; Transport kinetics