Histone Deacetylase Activity Assay

Part of the Methods in Molecular Biology book series (MIMB, volume 1288)

Abstract

Histone deacetylases (HDACs) are enzymes that catalyze the removal of acetyl groups from the ε-amino groups of conserved lysine residues in the amino terminal tail of histones. Accumulating evidence suggests that many, if not all, HDACs can also deacetylate nonhistone proteins. Through deacetylating histones and nonhistone proteins, HDACs regulate a variety of cellular processes including gene transcription, cell differentiation, DNA damage responses, and apoptosis. Aberrant HDACs are implicated in many human diseases and, therefore, it is important to have a consistent and reliable assay for analyzing HDAC activities. The focus of this chapter is to provide up-to-date, easy-to-follow, approaches and techniques, for the assay of HDAC enzymatic activities.

Key words

HDAC assay HDAC2 Sirtuin SIRT1 Protein deacetylation Core histone Boc-Lys(Ac)-AMC Radioactive assay Fluorometric assay 

References

  1. 1.
    Yang XJ, Seto E (2008) The Rpd3/Hda1 family of lysine deacetylases: from bacteria and yeast to mice and men. Nat Rev Mol Cell Biol 9(3):206–218CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Glozak MA, Sengupta N, Zhang X, Seto E (2005) Acetylation and deacetylation of non-histone proteins. Gene 363:15–23CrossRefPubMedGoogle Scholar
  3. 3.
    Peng L, Seto E (2011) Deacetylation of nonhistone proteins by HDACs and the implications in cancer. Handb Exp Pharmacol 206:39–56CrossRefPubMedGoogle Scholar
  4. 4.
    Carmen AA, Rundlett SE, Grunstein M (1996) HDA1 and HDA3 are components of a yeast histone deacetylase (HDA) complex. J Biol Chem 271(26):15837–15844CrossRefPubMedGoogle Scholar
  5. 5.
    Heltweg B, Jung M (2003) A homogeneous nonisotopic histone deacetylase activity assay. J Biomol Screen 8:89–95CrossRefPubMedGoogle Scholar
  6. 6.
    Hoffmann K et al (2000) First non-radioactive assay for in vitro screening of histone deacetylase inhibitors. Pharmazie 55:601–606PubMedGoogle Scholar
  7. 7.
    Wegener D et al (2003) A fluorogenic histone deacetylase assay well suited for high-throughput activity screening. Chem Biol 10:61–68CrossRefPubMedGoogle Scholar
  8. 8.
    Heltweg B, Trapp J, Jung M (2005) In vitro assays for the determination of histone deacetylase activity. Methods 36(4):332–337CrossRefPubMedGoogle Scholar
  9. 9.
    Rezai-Zadeh N, Tsai SC, Wen YD, Yao YL, Yang WM, Seto E (2004) Histone deacetylases: purification of the enzymes, substrates, and assay conditions. Methods Enzymol 377:167–179CrossRefPubMedGoogle Scholar
  10. 10.
    Dignam JD, Lebovitz RM, Roeder RG (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11:1475–1489CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Sun JM, Spencer VA, Chen HY, Li L, Davie JR (2003) Measurement of histone acetyltransferase and histone deacetylase activities and kinetics of histone acetylation. Methods 31(1):12–23CrossRefPubMedGoogle Scholar
  12. 12.
    Borra MT, Smith BC, Denu JM (2005) Mechanism of human SIRT1 activation by resveratrol. J Biol Chem 280(17):17187–17195CrossRefPubMedGoogle Scholar
  13. 13.
    Halley F, Reinshagen J, Ellinger B, Wolf M, Niles AL, Evans NJ, Kirkland TA, Wagner JM, Jung M, Gribbon P, Gul S (2011) A bioluminogenic HDAC activity assay: validation and screening. J Biomol Screen 16(10):1227–1235CrossRefPubMedGoogle Scholar
  14. 14.
    Lahm A, Paolini C, Pallaoro M, Nardi MC, Jones P, Neddermann P, Sambucini S, Bottomley MJ, Lo Surdo P, Carfí A, Koch U, De Francesco R, Steinkühler C, Gallinari P (2007) Unraveling the hidden catalytic activity of vertebrate class IIa histone deacetylases. Proc Natl Acad Sci U S A 104(44):17335–17340CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    Utley RT, Owen-Hughes TA, Juan LJ, Cote J, Adams CC, Workman JL (1996) In vitro analysis of transcription factor binding to nucleosomes and nucleosome disruption/displacement. Methods Enzymol 274:276–291CrossRefPubMedGoogle Scholar
  16. 16.
    Kolle D, Brosch G, Lechner T, Lusser A, Loidl P (1998) Biochemical methods for analysis of histone deacetylases. Methods 15(4):323–331CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Molecular Oncology DepartmentH. Lee Moffitt Cancer Center and Research InstituteTampaUSA
  2. 2.H. Lee Moffitt Cancer Center and Research InstituteTampaUSA

Personalised recommendations