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Nuclear Transport Receptor Binding Avidity Triggers A Self-healing Collapse Transition In FG-nucleoporin Molecular Brushes

Rafael L. Schoch, L. Kapinos, R. Y. Lim
Published 2012 · Chemistry, Medicine

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Conformational changes at supramolecular interfaces are fundamentally coupled to binding activity, yet it remains a challenge to probe this relationship directly. Within the nuclear pore complex, this underlies how transport receptors known as karyopherins proceed through a tethered layer of intrinsically disordered nucleoporin domains containing Phe-Gly (FG)-rich repeats (FG domains) that otherwise hinder passive transport. Here, we use nonspecific proteins (i.e., BSA) as innate molecular probes to explore FG domain conformational changes by surface plasmon resonance. This mathematically diminishes the surface plasmon resonance refractive index constraint, thereby providing the means to acquire and correlate height changes in a surface-tethered FG domain layer to Kap binding affinities in situ with respect to their relative spatial arrangements. Stepwise measurements show that FG domain collapse is caused by karyopherin β1 (Kapβ1) binding at low concentrations, but this gradually transitions into a reextension at higher Kapβ1 concentrations. This ability to self-heal is intimately coupled to Kapβ1-FG binding avidity that promotes the maximal incorporation of Kapβ1 into the FG domain layer. Further increasing Kapβ1 to physiological concentrations leads to a “pileup” of Kapβ1 molecules that bind weakly to unoccupied FG repeats at the top of the layer. Therefore, binding avidity does not hinder fast transport per se. Revealing the biophysical basis underlying the form–function relationship of Kapβ1-FG domain behavior results in a convergent picture in which transport and mechanistic aspects of nuclear pore complex functionality are reconciled.
This paper references
Structure and Function of Intrinsically Disordered Proteins (CRC, Boca Raton, FL)
P Tompa (2010)
10.1038/nnano.2011.88
Single-molecule transport across an individual biomimetic nuclear pore complex.
S. Kowalczyk (2011)
10.1021/nl801975q
Synthetic mimic of selective transport through the nuclear pore complex.
Yaron Caspi (2008)
10.1038/254109A0
Nuclear envelope permeability
P. L. Paine (1975)
10.1038/nmeth1171
Highly inclined thin illumination enables clear single-molecule imaging in cells
M. Tokunaga (2008)
10.1021/ac203096k
Multiparametric analysis of intrinsically disordered proteins: looking at intrinsic disorder through compound eyes.
V. Uversky (2012)
10.1021/BI012159+
Intrinsic Disorder and Protein Function
A. Dunker (2002)
10.1002/POLB.21203
Surface plasmon resonance as a tool for the estimation of adsorbed polymeric layer characteristics: Theoretical considerations and experiment
A. Koutsioubas (2007)
10.1002/bies.200800159
Translocation through the nuclear pore: Kaps pave the way
R. Peters (2009)
10.1038/nrm2114
Molecular mechanism of the nuclear protein import cycle
M. Stewart (2007)
10.1529/BIOPHYSJ.103.030072
The density and refractive index of adsorbing protein layers.
J. Vörös (2004)
10.1126/SCIENCE.1145980
Nanomechanical Basis of Selective Gating by the Nuclear Pore Complex
R. Y. Lim (2007)
10.1021/LA971228B
Quantitative interpretation of the response of surface plasmon resonance sensors to adsorbed films
Linda S. Jung (1998)
10.1103/PhysRevLett.89.238301
Morphology selection of nanoparticle dispersions by polymer media.
J. U. Kim (2002)
10.1083/jcb.200206106
Proteomic analysis of the mammalian nuclear pore complex
J. Cronshaw (2002)
10.4161/nucl.13818
'Natively unfolded' nucleoporins in nucleocytoplasmic transport
W. Yang (2011)
10.1016/J.COLSURFB.2007.09.029
My voyage of discovery to proteins in flatland ...and beyond.
W. Norde (2008)
10.1073/PNAS.0603521103
Flexible phenylalanine-glycine nucleoporins as entropic barriers to nucleocytoplasmic transport.
R. Y. Lim (2006)
10.1038/nmat2442
Understanding biophysicochemical interactions at the nano-bio interface.
A. Nel (2009)
10.1515/bc.2010.092
Converging on the function of intrinsically disordered nucleoporins in the nuclear pore complex
O. Peleg (2010)
10.1093/emboj/20.6.1320
Kinetic analysis of translocation through nuclear pore complexes
K. Ribbeck (2001)
10.1021/JP972635Z
Molecular Conformation in Oligo(ethylene glycol)-Terminated Self-Assembled Monolayers on Gold and Silver Surfaces Determines Their Ability To Resist Protein Adsorption
P. Harder (1998)
10.1074/mcp.M111.013656
Nucleocytoplasmic Transport: A Role for Nonspecific Competition in Karyopherin-Nucleoporin Interactions*
J. Tetenbaum-Novatt (2012)
10.1146/ANNUREV.BIOPHYS.26.1.541
Use of surface plasmon resonance to probe the equilibrium and dynamic aspects of interactions between biological macromolecules.
P. Schuck (1997)
10.1021/cr068107d
Surface plasmon resonance sensors for detection of chemical and biological species.
J. Homola (2008)
10.1016/0032-3861(77)90126-4
Van der Waals parameters, refractive indices and dispersion equations of spectrin, actin and other mammalian proteins
M. Andersen (1977)
10.1016/J.JMB.2006.09.089
Significant proportions of nuclear transport proteins with reduced intracellular mobilities resolved by fluorescence correlation spectroscopy.
Allison Paradise (2007)
10.1083/JCB.152.2.411
Gradient of Increasing Affinity of Importin β for Nucleoporins along the Pathway of Nuclear Import
I. Ben-Efraim (2001)
10.1083/jcb.200303085
Importin β contains a COOH-terminal nucleoporin binding region important for nuclear transport
Janna Bednenko (2003)
10.1074/mcp.M000035-MCP201
A Bimodal Distribution of Two Distinct Categories of Intrinsically Disordered Structures with Separate Functions in FG Nucleoporins*
J. Yamada (2010)
Quantitative structural analysis of importin-beta flexibility: Paradigm for Solenoid protein structures. Structure 18:1171–1183
JK Forwood (2010)
10.1021/JA00001A002
Formation of self-assembled monolayers by chemisorption of derivatives of oligo(ethylene glycol) of structure HS(CH2)11(OCH2CH2)mOH on gold
C. Pale-Grosdemange (1991)
10.1016/0925-4005(93)85239-7
Principles of biosensing with an extended coupling matrix and surface plasmon resonance
B. Liedberg (1993)
10.1038/nature06405
The molecular architecture of the nuclear pore complex
Frank Alber (2007)
10.1038/nmat2614
Emerging applications of stimuli-responsive polymer materials.
M. C. Stuart (2010)
10.4161/nucl.2.1.13818
'Natively unfolded' nucleoporins in nucleocytoplasmic transport: clustered or evenly distributed?
Weidong Yang (2011)
10.1073/pnas.1201724109
Self-regulated viscous channel in the nuclear pore complex
Jiong Ma (2012)
10.1073/pnas.0908269107
Three-dimensional distribution of transient interactions in the nuclear pore complex obtained from single-molecule snapshots
Jiong Ma (2010)
10.1364/JOSA.44.000362
Optical Constants of Silver, Gold, Copper, and Aluminum. II. The Index of Refraction n
L. Schulz (1954)
Formation of self - as - sembledmonolayers by chemisorption of derivatives of oligo ( ethylene glycol ) of structure Hs ( Ch 2 ) 11 ( Och 2 ch 2 ) Meta - Oh on Gold
C Palegrosdemange (1991)
10.1016/j.cell.2011.09.034
Molecular Architecture of the Transport Channel of the Nuclear Pore Complex
S. Solmaz (2011)
10.1016/j.cell.2007.06.024
A Saturated FG-Repeat Hydrogel Can Reproduce the Permeability Properties of Nuclear Pore Complexes
S. Frey (2007)
Quantitative Structural Analysis of Importin-β Flexibility: Paradigm for Solenoid Protein Structures
Jade K. Forwood (2010)
10.1371/journal.pcbi.0030125
Efficiency, Selectivity, and Robustness of Nucleocytoplasmic Transport
A. Zilman (2007)
10.1083/jcb.200605053
Nuclear import time and transport efficiency depend on importin β concentration
Weidong Yang (2006)
10.1016/0021-9797(91)90284-F
Quantitative determination of surface concentration of protein with surface plasmon resonance using radiolabeled proteins
E. Stenberg (1991)
10.1021/JA981897R
Using Self-Assembly To Control the Structure of DNA Monolayers on Gold: A Neutron Reflectivity Study
R. Levicky (1998)
10.1351/pac199567050829
Dextran modified self-assembled monolayer surfaces for use in biointeraction analysis with surface plasmon resonance
S. Lofas (1995)
10.1146/ANNUREV.PHYSCHEM.49.1.569
Interfaces and thin films as seen by bound electromagnetic waves.
W. Knoll (1998)
10.1016/S0092-8674(00)00014-3
Structural Basis for the Interaction between FxFG Nucleoporin Repeats and Importin-β in Nuclear Trafficking
R. Bayliss (2000)
10.1038/nature07600
Artificial nanopores that mimic the transport selectivity of the nuclear pore complex
T. Jovanović-Talisman (2009)
10.1016/J.SBI.2003.12.001
The interaction of proteins with solid surfaces.
J. J. Gray (2004)
10.1002/wnan.1160
Ordered and disordered proteins as nanomaterial building blocks.
N. Srinivasan (2012)
10.1083/JCB.148.4.635
The Yeast Nuclear Pore Complex: Composition, Architecture, and Transport Mechanism
M. Rout (2000)
10.1038/embor.2010.34
Ultrathin nucleoporin phenylalanine–glycine repeat films and their interaction with nuclear transport receptors
Nico B. Eisele (2010)
10.1038/emboj.2009.199
FG/FxFG as well as GLFG repeats form a selective permeability barrier with self-healing properties
S. Frey (2009)
10.1073/pnas.0437902100
Disorder in the nuclear pore complex: The FG repeat regions of nucleoporins are natively unfolded
D. Denning (2003)
10.1021/AC035326F
Evanescent field in surface plasmon resonance and surface plasmon field-enhanced fluorescence spectroscopies.
S. Ekgasit (2004)
Quantitative structural analysis of importin - beta flexibility : Paradigm for solenoid protein structures
JK Forwood (2010)
10.1364/JOSA.44.000357
The Optical Constants of Silver, Gold, Copper, and Aluminum. I. The Absorption Coefficient k
L. Schulz (1954)
10.1016/S0079-6107(01)00012-8
Predicting properties of intrinsically unstructured proteins.
J. Bright (2001)
10.1038/nrm1589
Intrinsically unstructured proteins and their functions
H. Dyson (2005)
10.1083/jcb.200204163
Influence of cargo size on Ran and energy requirements for nuclear protein import
S. K. Lyman (2002)
10.1038/nature10318
Nuclear export dynamics of RNA–protein complexes
D. Grünwald (2011)
10.1116/1.3407483
Protein resistant oligo(ethylene glycol) terminated self-assembled monolayers of thiols on gold by vapor deposition in vacuum
L. Kankate (2010)
10.1074/jbc.M109.095760
The Importin β Binding Domain Modulates the Avidity of Importin β for the Nuclear Pore Complex*
Kaylen Lott (2010)
10.1016/j.cell.2007.01.044
Natively Unfolded Nucleoporins Gate Protein Diffusion across the Nuclear Pore Complex
S. S. Patel (2007)
10.1016/J.STR.2005.09.007
Binding dynamics of isolated nucleoporin repeat regions to importin-beta.
Timothy A. Isgro (2005)
10.1146/annurev-biochem-060109-151030
The structure of the nuclear pore complex.
A. Hoelz (2011)



This paper is referenced by
10.1016/j.ceb.2017.05.003
The nuclear pore complex core scaffold and permeability barrier: variations of a common theme.
R. Hayama (2017)
10.1073/pnas.1212909110
Effect of charge, hydrophobicity, and sequence of nucleoporins on the translocation of model particles through the nuclear pore complex
M. Tagliazucchi (2013)
10.1021/acscentsci.8b00268
Gating Protein Transport in Solid State Nanopores by Single Molecule Recognition
Gustav Emilsson (2018)
10.3990/1.9789036540292
Size-selective analyte detection in an integrated optical Young interferometer biosensor
Harmen K P Mulder (2016)
10.1101/435651
Interactions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations
R. Hayama (2018)
10.1021/acs.biochem.8b00007
Structural Regulation of a Neurofilament-Inspired Intrinsically Disordered Protein Brush by Multisite Phosphorylation.
Ruoxing Lei (2018)
10.1007/978-3-319-77309-4_3
Dynamic Structures of the Nuclear Pore Complex and Their Roles in Nucleocytoplasmic Transport
M. Goldberg (2018)
10.1016/j.bpj.2014.07.060
Probing the disordered domain of the nuclear pore complex through coarse-grained molecular dynamics simulations.
Ali Ghavami (2014)
Dynamics of nuclear actin
Kari-Pekka Skarp (2014)
10.1021/acs.langmuir.6b01099
Site-Specific Modulation of Charge Controls the Structure and Stimulus Responsiveness of Intrinsically Disordered Peptide Brushes.
Maniraj Bhagawati (2016)
10.1021/acsami.5b01590
Strongly stretched protein resistant poly(ethylene glycol) brushes prepared by grafting-to.
Gustav Emilsson (2015)
10.1101/709675
Design principles of selective transport through biopolymer barriers
L. Maguire (2019)
10.1021/acs.langmuir.9b02538
Lipid Bilayer Thickness Measured by Quantitative DIC Reveals Phase Transitions and Effects of Substrate Hydrophilicity
D. Regan (2019)
10.1016/j.bpj.2013.09.006
Cohesiveness tunes assembly and morphology of FG nucleoporin domain meshworks - Implications for nuclear pore permeability.
Nico B. Eisele (2013)
TRÜPTOFAANI JA ARC-LUM(-) SONDI VAHELISE ENERGIAÜLEKANDE UURIMINE Bakalaurusetöö
Maikel Pulk (2014)
Coarse-grained molecular dynamics simulation of transport through the nuclear pore complex
Ali Ghavami (2014)
10.1021/ac403607s
Ion Permeability of the Nuclear Pore Complex and Ion-Induced Macromolecular Permeation as Studied by Scanning Electrochemical and Fluorescence Microscopy
J. Kim (2014)
10.1242/jcs.238121
Karyopherin enrichment at the nuclear pore complex attenuates Ran permeability
Suncica Barbato (2020)
10.1364/BOE.8.003606
Classification of biological micro-objects using optical coherence tomography: in silico study.
Paweł Ossowski (2017)
10.1063/1.4990796
Effects of cross-linking on partitioning of nanoparticles into a polymer brush: Coarse-grained simulations test simple approximate theories.
Masoumeh Ozmaian (2018)
10.1016/J.EML.2018.04.006
Sol-gel transition in solutions of FG-Nups of the nuclear pore complex
Ali Ghavami (2018)
JCB_201702092 3609..3624
Kapinos (2017)
10.1016/j.jmb.2016.01.002
The Multiple Faces of Disordered Nucleoporins.
E. Lemke (2016)
10.1016/j.str.2018.01.010
Deciphering the "Fuzzy" Interaction of FG Nucleoporins and Transport Factors Using Small-Angle Neutron Scattering.
Samuel Sparks (2018)
10.1093/jxb/eru346
Components of the Arabidopsis nuclear pore complex play multiple diverse roles in control of plant growth
G. Parry (2014)
Biophysical approach of neuronal shapes
Céline Braini (2016)
10.1083/jcb.201702092
Karyopherins regulate nuclear pore complex barrier and transport function
L. Kapinos (2017)
10.1016/j.apmt.2019.100478
A disposable microfluidic-integrated hand-held plasmonic platform for protein detection
F. Inci (2020)
10.7554/eLife.10785
Simple biophysics underpins collective conformations of the intrinsically disordered proteins of the Nuclear Pore Complex
A. Vovk (2016)
10.1371/journal.pcbi.1003488
Assembly of Nsp1 Nucleoporins Provides Insight into Nuclear Pore Complex Gating
Ramya Gamini (2014)
10.1103/physreve.101.022420
Intrinsically disordered nuclear pore proteins show ideal-polymer morphologies and dynamics.
L. K. Davis (2020)
10.1002/anie.201305359
Single-molecule analysis of the recognition forces underlying nucleo-cytoplasmic transport.
Martina Rangl (2013)
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