TY - JOUR
T1 - DNA origami scaffold for studying intrinsically disordered proteins of the nuclear pore complex
AU - Ketterer, Philip
AU - Ananth, Adithya N.
AU - Laman Trip, Diederik S.
AU - Mishra, Ankur
AU - Bertosin, Eva
AU - Ganji, Mahipal
AU - Van Der Torre, Jaco
AU - Onck, Patrick
AU - Dietz, Hendrik
AU - Dekker, Cees
PY - 2018/3/2
Y1 - 2018/3/2
N2 - 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.
AB - 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.
UR - http://resolverlink.tudelft.nl/uuid:ca1f7c4b-974f-4147-a7d7-a21644a494ec
UR - http://www.scopus.com/inward/record.url?scp=85042762674&partnerID=8YFLogxK
U2 - 10.1038/s41467-018-03313-w
DO - 10.1038/s41467-018-03313-w
M3 - Article
C2 - 29500415
SN - 2041-1723
VL - 9
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 902
ER -