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Analysis of Multivalent IDP Interactions: Stoichiometry, Affinity, and Local Concentration Effect Measurements

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Intrinsically Disordered Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2141))

Abstract

Nuclear magnetic resonance (NMR) titration and isothermal titration calorimetry can be combined to provide an assessment of how multivalent intrinsically disordered protein (IDP) interactions can involve enthalpy–entropy balance. Here, we describe the underlying technical details and additional methods, such as dynamic light scattering analysis, needed to assess these reactions. We apply this to a central interaction involving the disordered regions of phe–gly nucleoporins (FG-Nups) that contain multiple phenylalanine–glycine repeats which are of particular interest, as their interactions with nuclear transport factors (NTRs) underlie the paradoxically rapid yet also highly selective transport of macromolecules mediated by the nuclear pore complex (NPC). These analyses revealed that a combination of low per-FG motif affinity and the enthalpy–entropy balance prevents high-avidity interaction between FG-Nups and NTRs while the large number of FG motifs promotes frequent FG–NTR contacts, resulting in enhanced selectivity.

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Author information

Author notes

  1. Samuel Sparks

    Present address: Silicon Therapeutics, Boston, MA, USA

Authors and Affiliations

  1. Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY, USA

    Samuel Sparks & David Cowburn

  2. Laboratory of Cellular and Structural Biology, The Rockefeller University, New York, NY, USA

    Ryo Hayama & Michael P. Rout

Authors
  1. Samuel Sparks
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  2. Ryo Hayama
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  3. Michael P. Rout
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  4. David Cowburn
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Corresponding author

Correspondence to David Cowburn .

Editor information

Editors and Affiliations

  1. Structural Biology and NMR Laboratory, Department of Biology, University of Copenhagen, København N, Denmark

    Birthe B. Kragelund

  2. Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Copenhagen, Denmark

    Karen Skriver

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Sparks, S., Hayama, R., Rout, M.P., Cowburn, D. (2020). Analysis of Multivalent IDP Interactions: Stoichiometry, Affinity, and Local Concentration Effect Measurements. In: Kragelund, B.B., Skriver, K. (eds) Intrinsically Disordered Proteins. Methods in Molecular Biology, vol 2141. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0524-0_23

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  • DOI: https://doi.org/10.1007/978-1-0716-0524-0_23

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0523-3

  • Online ISBN: 978-1-0716-0524-0

  • eBook Packages: Springer Protocols

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