Frey, SteffenSteffenFreyRees, RenateRenateReesSchünemann, JürgenJürgenSchünemannNg, Sheung ChunSheung ChunNgFünfgeld, KevserKevserFünfgeldHuyton, TrevorTrevorHuytonGörlich, DirkDirkGörlich2022-07-042022-07-042018https://resolver.sub.uni-goettingen.de/purl?gro-2/112379Nuclear pore complexes (NPCs) conduct nucleocytoplasmic transport through an FG domain-controlled barrier. We now explore how surface-features of a mobile species determine its NPC passage rate. Negative charges and lysines impede passage. Hydrophobic residues, certain polar residues (Cys, His), and, surprisingly, charged arginines have striking translocation-promoting effects. Favorable cation-π interactions between arginines and FG-phenylalanines may explain this apparent paradox. Application of these principles to redesign the surface of GFP resulted in variants that show a wide span of transit rates, ranging from 35-fold slower than wild-type to ∼500 times faster, with the latter outpacing even naturally occurring nuclear transport receptors (NTRs). The structure of a fast and particularly FG-specific GFPNTR variant illustrates how NTRs can expose multiple regions for binding hydrophobic FG motifs while evading non-specific aggregation. Finally, we document that even for NTR-mediated transport, the surface-properties of the "passively carried" cargo can strikingly affect the translocation rate.enjournal_article10.1016/j.cell.2018.05.04529958108