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Recombinant Production of Monomeric Isotope-Enriched Aggregation-Prone Peptides: Polyglutamine Tracts and Beyond

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Intrinsically Disordered Proteins

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

Abstract

High solvent exposure of certain sequences located in intrinsically disordered regions (IDRs) may eventually lead to aggregation, as is the case for some low-complexity regions (LCRs) and short linear motifs (SLiMs). In particular, polyglutamine (polyQ) tracts are LCRs of variable length highly enriched in glutamine residues. They are common in transcription factors, and their length can have an impact on transcriptional activity. In nine proteins, polyQ tract expansions beyond specific thresholds cause nine neurodegenerative diseases, and aggregates formed by the protein harboring the polyQ tract can be detected in affected individuals. A structural characterization of polyQ proteins in their monomeric form is key to understand how their expansion can affect their aggregation propensity. In this regard, nuclear magnetic resonance (NMR) spectroscopy can provide high-resolution structural information. Here, we present a protocol to prepare monomeric samples of isotope-enriched short helical polyQ peptides based on the sequence of the androgen receptor (AR) suitable for NMR characterization and suggest different ways to adapt it for the production and monomerization of other relatively short IDR sequences and SLiMs.

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Author information

Author notes

  1. Giulio Chiesa

    Present address: Department of Biomedical Engineering and Biological Design Center, Boston University, Boston, MA, USA

Authors and Affiliations

  1. Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology, Barcelona, Spain

    Albert Escobedo, Giulio Chiesa & Xavier Salvatella

  2. Joint BSC-IRB Research Programme in Computational Biology, Barcelona, Spain

    Albert Escobedo, Giulio Chiesa & Xavier Salvatella

  3. ICREA, Barcelona, Spain

    Xavier Salvatella

Authors
  1. Albert Escobedo
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  2. Giulio Chiesa
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  3. Xavier Salvatella
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Corresponding author

Correspondence to Xavier Salvatella .

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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Escobedo, A., Chiesa, G., Salvatella, X. (2020). Recombinant Production of Monomeric Isotope-Enriched Aggregation-Prone Peptides: Polyglutamine Tracts and Beyond. 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_10

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  • DOI: https://doi.org/10.1007/978-1-0716-0524-0_10

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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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