DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex
Philip Ketterer (Technische Universität München)
Adithya Ananth (TU Delft - BN/Cees Dekker Lab, Kavli institute of nanoscience Delft)
D.S. Laman Trip (OLD BN/Hyun Youk Lab, Kavli institute of nanoscience Delft)
Ankur Mishra (University Medical Center Groningen)
Eva Bertosin (Technische Universität München)
M. Ganji (TU Delft - BN/Cees Dekker Lab, Kavli institute of nanoscience Delft)
J van der Torre (Kavli institute of nanoscience Delft, BN/Technici en Analisten)
Patrick Onck (University Medical Center Groningen)
Hendrik Dietz (Technische Universität München)
C Dekker (Kavli institute of nanoscience Delft, TU Delft - BN/Cees Dekker Lab)
More Info
expand_more
Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons.
Abstract
The nuclear pore complex (NPC) is the gatekeeper for nuclear transport in eukaryotic cells. A key component of the NPC is the central shaft lined with intrinsically disordered proteins (IDPs) known as FG-Nups, which control the selective molecular traffic. Here, we present an approach to realize artificial NPC mimics that allows controlling the type and copy number of FG-Nups. We constructed 34 nm-wide 3D DNA origami rings and attached different numbers of NSP1, a model yeast FG-Nup, or NSP1-S, a hydrophilic mutant. Using (cryo) electron microscopy, we find that NSP1 forms denser cohesive networks inside the ring compared to NSP1-S. Consistent with this, the measured ionic conductance is lower for NSP1 than for NSP1-S. Molecular dynamics simulations reveal spatially varying protein densities and conductances in good agreement with the experiments. Our technique provides an experimental platform for deciphering the collective behavior of IDPs with full control of their type and position.
help_outline