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Drug discovery assays for the histone deacetylase class of enzymes

18 December 2012  •  Author(s): Sheraz Gul and Gesa Witt, European ScreeningPort GmbH

The histone deacetylase (HDAC) class of enzyme are a group of conserved enzymes known for their ability to remove acetyl groups from lysine residues on histone tails. Since aberrant HDAC enzyme expression is observed in various diseases, there is increasing interest in finding small molecules which function as HDAC enzyme inhibitors. This article reviews the various biochemical assays available for monitoring HDAC enzyme activity that have been validated for use in High Throughput Screening. The assays referred to are compatible with standard microtitre plates (96 and 384 well format) and make use of absorbance, luminescence and fluorescence detection methods.

The histone deacetylase class of enzymes

The histone deacetylase (HDAC) class of enzymes are involved in many biological pathways1 and one of their best known properties is their ability to remove acetyl groups from lysine residues on amino-terminal histone tails2-4. Thus far, 18 HDAC enzymes have been identified which are divided into zinc dependent and NAD dependent enzymes5. The Class I HDAC enzymes include the zinc dependent HDACs 1, 2, 3, and 8 and consist of 350-500 amino acid residues6. The Class II HDAC enzymes are also zinc dependent but are larger, consist of about 1,000 amino acid residues7 and are subdivided into Class IIa (HDAC4, 5, 7, and 9) and Class IIb (HDAC6 and HDAC10) enzymes. The Class I and Class II HDAC enzymes can be inhibited by trichostatin A (TSA)8,9 and this inhibitor is often used as a reference to bench-mark their assays. The Class III HDAC enzymes are the sirtuin enzymes (SIRT1-7) and are NAD-dependent1,10. This class of enzymes is not sensitive to TSA but can be inhibited for example by nicotinamide.

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