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Structural Analyses of Intrinsically Disordered Proteins by Small-Angle X-Ray Scattering

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Intrinsically Disordered Proteins

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

Abstract

Small-angle X-ray scattering (SAXS) is a low-resolution method for the structural characterization of biological macromolecules in solution. Information about the overall structural features provided by SAXS is highly complementary to X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, and cryo-electron microscopy, which are high-resolution methods. SAXS not only provides the shape, oligomeric state, and quaternary structure of folded proteins and protein complexes but also allows for quantitative analysis of flexible biomolecules. In this chapter, the most relevant SAXS procedures for structural characterization of flexible macromolecules, including intrinsically disordered proteins (IDPs), are presented. The sample requirements for SAXS experiments on protein solutions and the sequence of steps in data collection and processing are described. The use of the advanced data analysis tools to quantitatively characterize flexible proteins is presented in detail. Typical experimental issues and potential problems encountered during SAXS data measurements and analyses are discussed.

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Acknowledgments

This work was supported by the Labex EpiGenMed, an “Investissements d’Avenir” program (ANR-10-LABX-12-01). The CBS is a member of France-BioImaging (FBI) and the French Infrastructure for Integrated Structural Biology (FRISBI), two national infrastructures supported by the French National Research Agency (ANR-10-INBS-04-01 and ANR-10-INBS-05, respectively). D.S. acknowledges support by iNEXT, grant number 653706, funded by the Horizon 2020 program of the European Union.

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Authors and Affiliations

  1. Centre de Biochimie Structurale, INSERM, CNRS, Université de Montpellier, Montpellier, France

    Amin Sagar & Pau Bernadó

  2. European Molecular Biology Laboratory, Hamburg Unit, EMBL c/o DESY, Hamburg, Germany

    Dmitri Svergun

Authors
  1. Amin Sagar
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  2. Dmitri Svergun
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  3. Pau Bernadó
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Corresponding authors

Correspondence to Amin Sagar or Pau Bernadó .

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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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Sagar, A., Svergun, D., Bernadó, P. (2020). Structural Analyses of Intrinsically Disordered Proteins by Small-Angle X-Ray Scattering. 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_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0524-0_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0523-3

  • Online ISBN: 978-1-0716-0524-0

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