skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Folding propensity of intrinsically disordered proteins by osmotic stress

Abstract

Proteins imparted with intrinsic disorder conduct a range of essential cellular functions. To better understand the folding and hydration properties of intrinsically disordered proteins (IDPs), we used osmotic stress to induce conformational changes in nuclear co-activator binding domain (NCBD) and activator for thyroid hormone and retinoid receptor (ACTR). Osmotic stress was applied by the addition of small and polymeric osmolytes, where we discovered that water contributions to NCBD folding always exceeded those for ACTR. Both NCBD and ACTR were found to gain a-helical structure with increasing osmotic stress, consistent with their folding upon NCBD/ACTR complex formation. Using small-angle neutron scattering (SANS), we further characterized NCBD structural changes with the osmolyte ethylene glycol. Here a large reduction in overall size initially occurred before substantial secondary structural change. In conclusion, by focusing on folding propensity, and linked hydration changes, we uncover new insights that may be important for how IDP folding contributes to binding.

Authors:
 [1];  [2];  [2];  [3];  [4];  [3];  [3];  [1];  [3]
  1. Univ. of Tennessee, Knoxville, TN (United States)
  2. Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  4. Univ. of Pittsburgh, Pittsburgh, PA (United States)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
Sponsoring Org.:
USDOE Laboratory Directed Research and Development (LDRD) Program
OSTI Identifier:
1333084
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Molecular BioSystems
Additional Journal Information:
Journal Volume: 12; Journal Issue: 12; Journal ID: ISSN 1742-206X
Publisher:
Royal Society of Chemistry
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Mansouri, Amanda L., Grese, Laura N., Rowe, Erica L., Pino, James C., Chennubhotla, S. Chakra, Ramanathan, Arvind, O'Neill, Hugh Michael, Berthelier, Valerie, and Stanley, Christopher B. Folding propensity of intrinsically disordered proteins by osmotic stress. United States: N. p., 2016. Web. doi:10.1039/C6MB00512H.
Mansouri, Amanda L., Grese, Laura N., Rowe, Erica L., Pino, James C., Chennubhotla, S. Chakra, Ramanathan, Arvind, O'Neill, Hugh Michael, Berthelier, Valerie, & Stanley, Christopher B. Folding propensity of intrinsically disordered proteins by osmotic stress. United States. doi:10.1039/C6MB00512H.
Mansouri, Amanda L., Grese, Laura N., Rowe, Erica L., Pino, James C., Chennubhotla, S. Chakra, Ramanathan, Arvind, O'Neill, Hugh Michael, Berthelier, Valerie, and Stanley, Christopher B. Tue . "Folding propensity of intrinsically disordered proteins by osmotic stress". United States. doi:10.1039/C6MB00512H. https://www.osti.gov/servlets/purl/1333084.
@article{osti_1333084,
title = {Folding propensity of intrinsically disordered proteins by osmotic stress},
author = {Mansouri, Amanda L. and Grese, Laura N. and Rowe, Erica L. and Pino, James C. and Chennubhotla, S. Chakra and Ramanathan, Arvind and O'Neill, Hugh Michael and Berthelier, Valerie and Stanley, Christopher B.},
abstractNote = {Proteins imparted with intrinsic disorder conduct a range of essential cellular functions. To better understand the folding and hydration properties of intrinsically disordered proteins (IDPs), we used osmotic stress to induce conformational changes in nuclear co-activator binding domain (NCBD) and activator for thyroid hormone and retinoid receptor (ACTR). Osmotic stress was applied by the addition of small and polymeric osmolytes, where we discovered that water contributions to NCBD folding always exceeded those for ACTR. Both NCBD and ACTR were found to gain a-helical structure with increasing osmotic stress, consistent with their folding upon NCBD/ACTR complex formation. Using small-angle neutron scattering (SANS), we further characterized NCBD structural changes with the osmolyte ethylene glycol. Here a large reduction in overall size initially occurred before substantial secondary structural change. In conclusion, by focusing on folding propensity, and linked hydration changes, we uncover new insights that may be important for how IDP folding contributes to binding.},
doi = {10.1039/C6MB00512H},
journal = {Molecular BioSystems},
number = 12,
volume = 12,
place = {United States},
year = {2016},
month = {10}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 1 work
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Mutual synergistic folding in recruitment of CBP/p300 by p160 nuclear receptor coactivators
journal, January 2002

  • Demarest, Stephen J.; Martinez-Yamout, Maria; Chung, John
  • Nature, Vol. 415, Issue 6871
  • DOI: 10.1038/415549a

Helical Propensity in an Intrinsically Disordered Protein Accelerates Ligand Binding
journal, January 2014

  • Iešmantavičius, Vytautas; Dogan, Jakob; Jemth, Per
  • Angewandte Chemie International Edition, Vol. 53, Issue 6
  • DOI: 10.1002/anie.201307712

Conformations of intrinsically disordered proteins are influenced by linear sequence distributions of oppositely charged residues
journal, July 2013

  • Das, R. K.; Pappu, R. V.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 33
  • DOI: 10.1073/pnas.1304749110

Mantid—Data analysis and visualization package for neutron scattering and μ SR experiments
journal, November 2014

  • Arnold, O.; Bilheux, J. C.; Borreguero, J. M.
  • Nuclear Instruments and Methods in Physics Research Section A: Accelerators, Spectrometers, Detectors and Associated Equipment, Vol. 764
  • DOI: 10.1016/j.nima.2014.07.029

A decade and a half of protein intrinsic disorder: Biology still waits for physics: Protein Intrinsic Disorder
journal, April 2013


Conformational selection in the molten globule state of the nuclear coactivator binding domain of CBP
journal, June 2010

  • Kjaergaard, Magnus; Teilum, Kaare; Poulsen, Flemming M.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 28
  • DOI: 10.1073/pnas.1001693107

Structural thermodynamics of protein preferential solvation: Osmolyte solvation of proteins, aminoacids, and peptides
journal, May 2008

  • Auton, Matthew; Bolen, D. Wayne; Rösgen, Jörg
  • Proteins: Structure, Function, and Bioinformatics, Vol. 73, Issue 4
  • DOI: 10.1002/prot.22103

Differences in Stability among the Human Apolipoprotein E Isoforms Determined by the Amino-Terminal Domain
journal, September 2000

  • Morrow, Julie A.; Segall, Mark L.; Lund-Katz, Sissel
  • Biochemistry, Vol. 39, Issue 38
  • DOI: 10.1021/bi000099m

Absolute calibration of small-angle neutron scattering data
journal, February 1987


NMR Relaxation Study of the Complex Formed Between CBP and the Activation Domain of the Nuclear Hormone Receptor Coactivator ACTR
journal, February 2008

  • Ebert, Marc-Olivier; Bae, Sung-Hun; Dyson, H. Jane
  • Biochemistry, Vol. 47, Issue 5
  • DOI: 10.1021/bi701767j

Forced Folding of a Disordered Protein Accesses an Alternative Folding Landscape
journal, October 2014

  • Moosa, Mahdi Muhammad; Ferreon, Allan Chris M.; Deniz, Ashok A.
  • ChemPhysChem, Vol. 16, Issue 1
  • DOI: 10.1002/cphc.201402661

Determination of the regularization parameter in indirect-transform methods using perceptual criteria
journal, August 1992


Minimal Effects of Macromolecular Crowding on an Intrinsically Disordered Protein: A Small-Angle Neutron Scattering Study
journal, February 2014


Fast Association and Slow Transitions in the Interaction between Two Intrinsically Disordered Protein Domains
journal, August 2012

  • Dogan, Jakob; Schmidt, Tanja; Mu, Xin
  • Journal of Biological Chemistry, Vol. 287, Issue 41
  • DOI: 10.1074/jbc.M112.399436

Modulation of the Intrinsic Helix Propensity of an Intrinsically Disordered Protein Reveals Long-Range Helix–Helix Interactions
journal, June 2013

  • Iešmantavičius, Vytautas; Jensen, Malene Ringkjøbing; Ozenne, Valéry
  • Journal of the American Chemical Society, Vol. 135, Issue 27
  • DOI: 10.1021/ja4045532

Measuring Osmotic Pressure of Poly(ethylene glycol) Solutions by Sedimentation Equilibrium Ultracentrifugation
journal, September 2003

  • Stanley, Christopher B.; Strey, Helmut H.
  • Macromolecules, Vol. 36, Issue 18
  • DOI: 10.1021/ma034079e

Intrinsically disordered proteins in cellular signalling and regulation
journal, December 2014

  • Wright, Peter E.; Dyson, H. Jane
  • Nature Reviews Molecular Cell Biology, Vol. 16, Issue 1
  • DOI: 10.1038/nrm3920

Linking folding and binding
journal, February 2009


Trimethylamine N -Oxide-induced Cooperative Folding of an Intrinsically Unfolded Transcription-activating Fragment of Human Glucocorticoid Receptor
journal, April 1999

  • Baskakov, Ilia V.; Kumar, Raj; Srinivasan, Ganesan
  • Journal of Biological Chemistry, Vol. 274, Issue 16
  • DOI: 10.1074/jbc.274.16.10693

Quantitative Biophysical Characterization of Intrinsically Disordered Proteins
journal, February 2015

  • Gibbs, Eric B.; Showalter, Scott A.
  • Biochemistry, Vol. 54, Issue 6
  • DOI: 10.1021/bi501460a

Protein Disorder Prevails under Crowded Conditions
journal, July 2011

  • Szasz, Cs.; Alexa, A.; Toth, K.
  • Biochemistry, Vol. 50, Issue 26
  • DOI: 10.1021/bi200365j

Effects of Macromolecular Crowding on the Intrinsically Disordered Proteins c-Fos and p27 Kip1
journal, June 2001

  • Flaugh, Shannon L.; Lumb, Kevin J.
  • Biomacromolecules, Vol. 2, Issue 2
  • DOI: 10.1021/bm015502z

MULCh : modules for the analysis of small-angle neutron contrast variation data from biomolecular assemblies
journal, January 2008

  • Whitten, Andrew E.; Cai, Shuzhi; Trewhella, Jill
  • Journal of Applied Crystallography, Vol. 41, Issue 1
  • DOI: 10.1107/S0021889807055136

Enthalpically driven peptide stabilization by protective osmolytes
journal, January 2010

  • Politi, Regina; Harries, Daniel
  • Chemical Communications, Vol. 46, Issue 35
  • DOI: 10.1039/c0cc01763a

Unusual biophysics of intrinsically disordered proteins
journal, May 2013


JPred4: a protein secondary structure prediction server
journal, April 2015

  • Drozdetskiy, Alexey; Cole, Christian; Procter, James
  • Nucleic Acids Research, Vol. 43, Issue W1
  • DOI: 10.1093/nar/gkv332

Classification of Intrinsically Disordered Regions and Proteins
journal, December 2013

  • van der Lee, Robin; Buljan, Marija; Lang, Benjamin
  • Chemical Reviews, Vol. 114, Issue 13
  • DOI: 10.1021/cr400525m

Effects of Macromolecular Crowding on the Conformational Ensembles of Disordered Proteins
journal, September 2013

  • Qin, Sanbo; Zhou, Huan-Xiang
  • The Journal of Physical Chemistry Letters, Vol. 4, Issue 20
  • DOI: 10.1021/jz401817x

A Folded Excited State of Ligand-Free Nuclear Coactivator Binding Domain (NCBD) Underlies Plasticity in Ligand Recognition
journal, February 2013

  • Kjaergaard, Magnus; Andersen, Lisbeth; Nielsen, Lau Dalby
  • Biochemistry, Vol. 52, Issue 10
  • DOI: 10.1021/bi4001062

The extended Q -range small-angle neutron scattering diffractometer at the SNS
journal, July 2010


Balance of enthalpy and entropy in depletion forces
journal, December 2013

  • Sukenik, Shahar; Sapir, Liel; Harries, Daniel
  • Current Opinion in Colloid & Interface Science, Vol. 18, Issue 6
  • DOI: 10.1016/j.cocis.2013.10.002

Osmolyte-Induced Folding of an Intrinsically Disordered Protein: Folding Mechanism in the Absence of Ligand
journal, June 2010

  • Chang, Yu-Chu; Oas, Terrence G.
  • Biochemistry, Vol. 49, Issue 25
  • DOI: 10.1021/bi100222h

Solvent effects in the helix-coil transition model can explain the unusual biophysics of intrinsically disordered proteins
journal, July 2015

  • Badasyan, Artem; Mamasakhlisov, Yevgeni Sh.; Podgornik, Rudolf
  • The Journal of Chemical Physics, Vol. 143, Issue 1
  • DOI: 10.1063/1.4923292

Macromolecular Crowding Effects on Coupled Folding and Binding
journal, October 2014

  • Kim, Young C.; Bhattacharya, Apratim; Mittal, Jeetain
  • The Journal of Physical Chemistry B, Vol. 118, Issue 44
  • DOI: 10.1021/jp508046y

Mapping Unstructured Regions and Synergistic Folding in Intrinsically Disordered Proteins with Amide H/D Exchange Mass Spectrometry
journal, October 2011

  • Keppel, Theodore R.; Howard, Brent A.; Weis, David D.
  • Biochemistry, Vol. 50, Issue 40
  • DOI: 10.1021/bi200875p

Exploring Free-Energy Landscapes of Intrinsically Disordered Proteins at Atomic Resolution Using NMR Spectroscopy
journal, December 2013

  • Jensen, Malene Ringkjøbing; Zweckstetter, Markus; Huang, Jie-rong
  • Chemical Reviews, Vol. 114, Issue 13
  • DOI: 10.1021/cr400688u

The transition state structure for coupled binding and folding of disordered protein domains
journal, June 2013

  • Dogan, Jakob; Mu, Xin; Engström, Åke
  • Scientific Reports, Vol. 3, Issue 1
  • DOI: 10.1038/srep02076

Single-molecule spectroscopy reveals polymer effects of disordered proteins in crowded environments
journal, March 2014

  • Soranno, Andrea; Koenig, Iwo; Borgia, Madeleine B.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 13
  • DOI: 10.1073/pnas.1322611111