Open AccessInteractions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations
Author(s) -
Ryo Hayama,
Mirco Sorci,
John J. Keating,
Lee M. Hecht,
Joel L. Plawsky,
Georges Belfort,
Brian T. Chait,
Michael P. Rout
Publication year - 2019
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0217897
Subject(s) - nucleoporin , nuclear pore , quartz crystal microbalance , biophysics , surface plasmon resonance , protein–protein interaction , nuclear transport , plasma protein binding , chemistry , transport protein , chemical physics , materials science , nanotechnology , microbiology and biotechnology , biology , nucleus , cell nucleus , nanoparticle , biochemistry , organic chemistry , adsorption
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.