Chapter title |
Histone Deacetylases
|
---|---|
Chapter number | 8 |
Book title |
Histone Deacetylases
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3667-0_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3665-6, 978-1-4939-3667-0
|
Authors |
Marek, Martin, Shaik, Tajith B, Duclaud, Sylvie, Pierce, Raymond J, Romier, Christophe, Martin Marek, Tajith B. Shaik, Sylvie Duclaud, Raymond J. Pierce, Christophe Romier, Shaik, Tajith B., Pierce, Raymond J. |
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. |
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