title: Structural characterization of nanoscale meshworks within a nucleoporin FG hydrogel.
abstract: The permeability barrier of nuclear pore complexes (NPCs) controls all exchange of macromolecules between the cytoplasm and the cell nucleus. It consists of phenylalanine-glycine (FG) repeat domains apparently organized as an FG hydrogel. It has previously been demonstrated that an FG hydrogel derived from the yeast nucleoporin Nsp1p reproduces the selectivity of authentic NPCs. Here we combined time-resolved optical spectroscopy and X-ray scattering techniques to characterize such a gel. The data suggest a hierarchy of structures that form during gelation at the expense of unstructured elements. On the largest scale, protein-rich domains with a correlation length of ~16.5 nm are evident. On a smaller length scale, aqueous channels with an average diameter of ~3 nm have been found, which possibly represent the physical structures accounting for the passive sieving effect of nuclear pores. The protein-rich domains contain characteristic β-structures with typical inter-β-strand and inter-β-sheet distances of 1.3 and 0.47 nm, respectively. During gelation, the formation of oligomeric associates is accompanied by the transfer of phenylalanines into a hydrophobic microenvironment, supporting the view that this process is driven by a hydrophobic collapse.
authors.affiliation: Research Group of Structural Dynamics of (Bio)chemical Systems, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.
authors.forename: Marcel
authors.initials: M
authors.lastname: Petri
authors.forename: Steffen
authors.initials: S
authors.lastname: Frey
authors.forename: Andreas
authors.initials: A
authors.lastname: Menzel
authors.forename: Dirk
authors.initials: D
authors.lastname: Görlich
authors.forename: Simone
authors.initials: S
authors.lastname: Techert
mesh_terms: D005998:Glycine / Q000737:chemistry*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D046911:Macromolecular Substances / Q000737:chemistry; D049329:Nanostructures / Q000737:chemistry*; D010649:Phenylalanine / Q000737:chemistry*; D011487:Protein Conformation
chemical_list: D046911:Macromolecular Substances; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D010649:Phenylalanine; D005998:Glycine