title: How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot.
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).
authors.affiliation: Institute of Toxicology, University Medical Center Mainz, Mainz, 55131, Germany.
authors.forename: Mandy
authors.initials: M
authors.lastname: Beyer
authors.affiliation: Institute of Toxicology, University Medical Center Mainz, Mainz, 55131, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Kiweler
authors.affiliation: Faculty of Chemistry and Pharmacy, Institute of Pharmacy, University of Regensburg, Regensburg, 93040, Germany.
authors.forename: Siavosh
authors.initials: S
authors.lastname: Mahboobi
authors.affiliation: Institute of Toxicology, University Medical Center Mainz, Mainz, 55131, Germany. okraemer@uni-mainz.de.
authors.forename: Oliver H
authors.initials: OH
authors.lastname: Krämer
mesh_terms: D000107:Acetylation; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D044127:Epigenesis, Genetic; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D056284:Histone Deacetylase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D056464:Histone Deacetylase 2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000073864:Histone Deacetylase 6 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D051379:Mice; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D000077767:Panobinostat; D011499:Protein Processing, Post-Translational; D011725:Pyridines / Q000494:pharmacology; D014635:Valproic Acid / Q000494:pharmacology; D000099186:Histone Deacetylase 3
chemical_list: D001549:Benzamides; D056284:Histone Deacetylase 1; D056464:Histone Deacetylase 2; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases; D006657:Histones; D006877:Hydroxamic Acids; D007211:Indoles; D000077767:Panobinostat; D011725:Pyridines; D014635:Valproic Acid; D000099186:Histone Deacetylase 3; C118739:entinostat; C110550:HDAC1 protein, human; C533423:HDAC2 protein, human; C433811:HDAC6 protein, human
keywords: Acetylation; HDAC6; HDACi; HDACs; Histone; Tubulin