title: Structural analysis of the p62 complex, an assembly of O-linked glycoproteins that localizes near the central gated channel of the nuclear pore complex.
abstract: The p62 complex is an oligomeric assembly of O-linked glycoproteins of the nuclear pore complex that interacts with cytosolic transport factors and is part of the machinery for nuclear protein import. In this study we have purified the p62 complex from rat liver nuclear envelopes and analyzed its structure and composition. The p62 complex consists of four distinct polypeptides (p62, p58, p54, and p45) and has a mass of approximately 234 kDa, calculated from its hydrodynamic properties and supported by chemical cross-linking and scanning transmission electron microscopy. These data suggest that the p62 complex contains one copy of each constituent polypeptide. Analysis of preparations of the p62 complex by electron microscopy using rotary metal shadowing and negative staining revealed donut-shaped particles with a diameter of approximately 15 nm. Immunogold electron microscopy of isolated rat liver nuclear envelopes demonstrated that p62 occurs on both the nucleoplasmic and cytoplasmic sides of the pore complex near the central gated channel involved in active transport of proteins and RNAs. The properties and localization of the p62 complex suggest that it may be involved in binding transport ligands near the center of the nuclear pore complex and in subsequently transferring them to the gated transport channel.
authors.affiliation: Department of Cell and Molecular Biology, Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: T
authors.initials: T
authors.lastname: Guan
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.forename: G
authors.initials: G
authors.lastname: Klier
authors.forename: N
authors.initials: N
authors.lastname: Panté
authors.forename: J M
authors.initials: JM
authors.lastname: Blevitt
authors.forename: M
authors.initials: M
authors.lastname: Haner
authors.forename: B
authors.initials: B
authors.lastname: Paschal
authors.forename: U
authors.initials: U
authors.lastname: Aebi
authors.forename: L
authors.initials: L
authors.lastname: Gerace
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D007473:Ion Channels / Q000502:physiology / Q000648:ultrastructure*; D008099:Liver / Q000648:ultrastructure; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*; D017348:Microscopy, Electron, Scanning Transmission; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D009685:Nuclear Envelope / Q000502:physiology / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure*; D051381:Rats
chemical_list: D007473:Ion Channels; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D009687:Nuclear Proteins