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Large-Scale Overproduction and Purification of Recombinant Histone Deacetylase 8 (HDAC8) from the Human-Pathogenic Flatworm Schistosoma mansoni

  • Martin Marek
  • Tajith B. Shaik
  • Sylvie Duclaud
  • Raymond J. Pierce
  • Christophe RomierEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1436)

Abstract

Epigenetic mechanisms underlie the morphological transformations and shifts in virulence of eukaryotic pathogens. The targeting of epigenetics-driven cellular programs thus represents an Achilles’ heel of human parasites. Today, zinc-dependent histone deacetylases (HDACs) belong to the most explored epigenetic drug targets in eukaryotic parasites. Here, we describe an optimized protocol for the large-scale overproduction and purification of recombinant smHDAC8, an emerging epigenetic drug target in the multicellular human-pathogenic flatworm Schistosoma mansoni. The strategy employs the robustness of recombinant expression in Escherichia coli together with initial purification through a poly-histidine affinity tag that can be removed by the thrombin protease. This protocol is divided into two steps: (1) large-scale production of smHDAC8 in E. coli, and (2) purification of the target smHDAC8 protein through multiple purification steps.

Key words

Histone deacetylase Enzyme Recombinant expression Purification Schistosoma 

Notes

Acknowledgements

This work and the authors of this manuscript have been supported by funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreements nos. 241865 (SEtTReND) and 602080 (A-ParaDDisE). The authors are supported by institutional funds from the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM), the Université de Strasbourg and the Université de Lille 2, the French Infrastructure for Integrated Structural Biology (FRISBI; ANR-10-INSB-05-01), and by Instruct as part of the European Strategy Forum on Research Infrastructures (ESFRI).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Martin Marek
    • 1
  • Tajith B. Shaik
    • 1
  • Sylvie Duclaud
    • 1
  • Raymond J. Pierce
    • 2
  • Christophe Romier
    • 1
    Email author
  1. 1.Département de Biologie Structurale Intégrative, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC)Université de Strasbourg (UDS), CNRS, INSERMIllkirch CedexFrance
  2. 2.Center for Infection and Immunity of Lille (CIIL), INSERM U1019 – CNRS UMR 8204Université de Lille, Institut Pasteur de LilleLille CedexFrance

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