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Assessment of HDACi-Induced Protein Cleavage by Caspases

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Book cover HDAC/HAT Function Assessment and Inhibitor Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1510))

Abstract

Aberrant histone deacetylase (HDAC) activity often correlates with neoplastic transformation and inhibition of HDACs by small molecules has emerged as a promising strategy to treat hematological malignancies in particular. Treatment with HDAC inhibitors (HDACis) often prompts tumor cells to undergo apoptosis, thereby causing a caspase-dependent cleavage of target proteins. An unexpectedly large number of proteins are in vivo caspase substrates and defining caspase-mediated substrate specificity is a major challenge. In this chapter we demonstrate that the hematopoietic transcription factor PU.1 becomes cleaved after treatment of acute myeloid leukemia (AML) cells with the HDACis LBH589 (panobinostat) or MS-275 (entinostat). To define caspase specificity for PU.1, an in vitro caspase assay including caspases 1–10 with in vitro-translated PU.1 is described in detail.

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Acknowledgements

This work was supported by the START program of the Medical School of the Rheinische-Westfälische Technische Hochschule (RWTH) Aachen University to J. Hartkamp.

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Authors and Affiliations

  1. Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany

    Fabian Treude, Tobias Gladbach, Jacqueline Plaster & Jörg Hartkamp

Authors
  1. Fabian Treude
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  2. Tobias Gladbach
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  3. Jacqueline Plaster
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  4. Jörg Hartkamp
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Corresponding author

Correspondence to Jörg Hartkamp .

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Editors and Affiliations

  1. Institute of Toxicology, University Medical Center Mainz, Mainz, Germany

    Oliver H. Krämer

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Treude, F., Gladbach, T., Plaster, J., Hartkamp, J. (2017). Assessment of HDACi-Induced Protein Cleavage by Caspases. In: Krämer, O. (eds) HDAC/HAT Function Assessment and Inhibitor Development. Methods in Molecular Biology, vol 1510. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6527-4_2

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  • DOI: https://doi.org/10.1007/978-1-4939-6527-4_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6525-0

  • Online ISBN: 978-1-4939-6527-4

  • eBook Packages: Springer Protocols

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