Unstable Protein Purification Through the Formation of Stable Complexes

  • Sylvia Eiler
  • Nicolas Levy
  • Benoit Maillot
  • Julien Batisse
  • Karine Pradeau Aubreton
  • Oyindamola Oladosu
  • Marc Ruff
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1764)

Abstract

Purification of proteins containing disordered regions and participating in transient complexes is often challenging because of the small amounts available after purification, their heterogeneity, instability, and/or poor solubility. To circumvent these difficulties, we set up a methodology that enables the production of stable complexes in large amounts for structural and functional studies. In this chapter, we describe the methodology used to establish the best cell culture conditions and buffer compositions to optimize soluble protein production and their stabilization through protein complex formation. Two examples of challenging protein families are described, namely, the human steroid nuclear receptors and the HIV-1 pre-integration complexes.

Key words

Protein complex Nuclear receptor GR TIF2 HIV Integrase Pre-integration complex 

Notes

Acknowledgments

This work was supported by grants from the CNRS, the INSERM, SIDACTION, and the French National Agency for Research against AIDS (ANRS), the support and the use of resources of the French Infrastructure for Integrated Structural Biology (FRISBI) ANR-10-INSB-05 and of Instruct, a Landmark ESFRI project. We wish to thank Robert Drillien (IGBMC) for his help and for useful suggestions about the manuscript. We would like to thank the members of the IGBMC Structural Biology and Genomics platform, the IGBMC cloning service headed by Paola Rossolillo, and the members of the IGBMC common services for their contribution.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Sylvia Eiler
    • 1
  • Nicolas Levy
    • 1
  • Benoit Maillot
    • 1
  • Julien Batisse
    • 1
  • Karine Pradeau Aubreton
    • 1
  • Oyindamola Oladosu
    • 1
  • Marc Ruff
    • 1
  1. 1.IGBMCIllkirchFrance

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