How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot

Methods Mol Biol. 2017:1510:355-364. doi: 10.1007/978-1-4939-6527-4_26.

Abstract

Histone deacetylases (HDACs) catalyze the deacetylation of lysine residues in their target proteins. This biochemical modification can have profound effects on the functions of these proteins and a dysregulation of HDAC activity contributes to severe diseases, including neoplastic transformation. In the following chapter, we present a strategy that allows to distinguish between the inhibition of the class I HDACs HDAC1, 2, and 3 and of the class IIb HDAC HDAC6. This method is based on Western blot and relies on the detection of hyperacetylated substrates of class I or class IIb HDACs in lysates from cells that were treated with histone deacetylase inhibitors (HDACi).

Keywords: Acetylation; HDAC6; HDACi; HDACs; Histone; Tubulin.

MeSH terms

  • Acetylation
  • Animals
  • Benzamides / pharmacology
  • Blotting, Western / methods
  • Cell Line, Tumor
  • Epigenesis, Genetic
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / enzymology
  • Histone Deacetylase 1 / antagonists & inhibitors
  • Histone Deacetylase 1 / genetics
  • Histone Deacetylase 1 / metabolism*
  • Histone Deacetylase 2 / antagonists & inhibitors
  • Histone Deacetylase 2 / genetics
  • Histone Deacetylase 2 / metabolism*
  • Histone Deacetylase 6 / antagonists & inhibitors
  • Histone Deacetylase 6 / genetics
  • Histone Deacetylase 6 / metabolism*
  • Histone Deacetylase Inhibitors / pharmacology*
  • Histone Deacetylases / genetics
  • Histone Deacetylases / metabolism*
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Hydroxamic Acids / pharmacology
  • Indoles / pharmacology
  • Mice
  • Monocytes / cytology
  • Monocytes / drug effects
  • Monocytes / enzymology
  • Panobinostat
  • Protein Processing, Post-Translational*
  • Pyridines / pharmacology
  • Valproic Acid / pharmacology

Substances

  • Benzamides
  • Histone Deacetylase Inhibitors
  • Histones
  • Hydroxamic Acids
  • Indoles
  • Pyridines
  • entinostat
  • Valproic Acid
  • Panobinostat
  • HDAC1 protein, human
  • HDAC2 protein, human
  • HDAC6 protein, human
  • Histone Deacetylase 1
  • Histone Deacetylase 2
  • Histone Deacetylase 6
  • Histone Deacetylases
  • histone deacetylase 3