Interactions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations

PLoS One. 2019 Jun 6;14(6):e0217897. doi: 10.1371/journal.pone.0217897. eCollection 2019.

Abstract

Protein-protein interactions are central to biological processes. In vitro methods to examine protein-protein interactions are generally categorized into two classes: in-solution and surface-based methods. Here, using the multivalent interactions between nucleocytoplasmic transport factors and intrinsically disordered FG repeat containing nuclear pore complex proteins as a model system, we examined the utility of three surface-based methods: atomic force microscopy, quartz crystal microbalance with dissipation, and surface plasmon resonance. Although results were comparable to those of previous reports, the apparent effect of mass transport limitations was demonstrated. Additional experiments with a loss-of-interaction FG repeat mutant variant demonstrated that the binding events that take place on surfaces can be unexpectedly complex, suggesting particular care must be exercised in interpretation of such data.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Active Transport, Cell Nucleus
  • Amino Acid Sequence
  • Cell Nucleus / metabolism*
  • Mutation / genetics
  • Nuclear Pore Complex Proteins / chemistry*
  • Nuclear Pore Complex Proteins / metabolism*
  • Protein Binding
  • Quartz Crystal Microbalance Techniques
  • Repetitive Sequences, Nucleic Acid*
  • beta Karyopherins / metabolism

Substances

  • Nuclear Pore Complex Proteins
  • beta Karyopherins