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Production of Intrinsically Disordered Proteins for Biophysical Studies: Tips and Tricks

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Intrinsically Disordered Proteins

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

Abstract

Intrinsically disordered proteins (IDPs) have no single, fixed tertiary structure, yet they take on many vital functions in biology. In recent years, considerable effort has been put into the structural characterization of their conformational ensembles, to understand the link between the transient, short- and long-range organizations of IDPs and their functions. Such biophysical studies require substantial amounts of pure protein, representing a major bottleneck in the studies of IDPs. However, the unique physicochemical properties resulting from their compositional bias may be exploited for simple yet effective purification strategies. In this chapter, we provide tips and tricks for IDP production and describe the most important analyses to carry out before bringing an IDP of interest to the laboratory. We outline four purification protocols utilizing the unique properties of IDPs as well as some commonly encountered challenges and pitfalls.

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Acknowledgments

We are grateful to the members of SBiNLab for many insightful discussions on IDPs and purification. This work is a contribution from the Novo Nordisk Foundation Challenge Center REPIN.

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Author notes

  1. Christian Parsbæk Pedersen and Pernille Seiffert contributed equally to this work.

Authors and Affiliations

  1. Structural Biology and NMR Laboratory and the Linderstrøm-Lang Centre for Protein Science, Department of Biology, University of Copenhagen, Copenhagen N, Denmark

    Christian Parsbæk Pedersen, Pernille Seiffert, Inna Brakti & Katrine Bugge

Authors
  1. Christian Parsbæk Pedersen
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  2. Pernille Seiffert
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  3. Inna Brakti
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  4. Katrine Bugge
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Corresponding author

Correspondence to Katrine Bugge .

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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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Pedersen, C.P., Seiffert, P., Brakti, I., Bugge, K. (2020). Production of Intrinsically Disordered Proteins for Biophysical Studies: Tips and Tricks. 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_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0524-0_9

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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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