Measurement and Analysis of Lysine Acetylation by KAT Complexes In Vitro and In Vivo

Part of the Methods in Molecular Biology book series (MIMB, volume 1983)


The acetylation of the ε-amine of lysine residues has significant impacts on the cellular functions of proteins. Through the combination of unbiased and targeted analysis of acetylated proteins, biological insights on lysine acetylation are now routinely generated. To help in this endeavor, we describe detailed protocols for the identification of acetylated lysine residues and the preparation of multiple reagents for the characterization of these sites in order to obtain functional insights on this widespread modification.


Lysine acetylation KAT HAT TAP LC-MS/MS 



We are grateful to Yannick Doyon, Karine Jacquet, Pierre Billon, and Xue Cheng for sharing their protocols. Research in the Côté and Lambert laboratories is funded by a Discovery Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC) (1304616-2017 to J.-P.L.) and a Foundation Grant from the Canadian Institutes of Health Research (CIHR) (FDN-143314 to J.C.). J.-P.L. is supported by a Junior 1 salary award from the Fonds de Recherche du Québec-Santé (FRQ-S; 251747), by an Operating Grant from the Cancer Research Society (22779) and a Leader’s Opportunity Funds from the Canada Foundation for Innovation (37454). J.C. holds a Canada Research Chair (Tier 1) in Chromatin Biology and Molecular Epigenetics.


  1. 1.
    Kouzarides T (2000) Acetylation: a regulatory modification to rival phosphorylation? EMBO J 19(6):1176–1179CrossRefGoogle Scholar
  2. 2.
    Allfrey VG, Mirsky AE (1964) Structural modifications of histones and their possible role in the regulation of RNA synthesis. Science 144(3618):559CrossRefGoogle Scholar
  3. 3.
    Pogo BG, Allfrey VG, Mirsky AE (1966) RNA synthesis and histone acetylation during the course of gene activation in lymphocytes. Proc Natl Acad Sci U S A 55(4):805–812CrossRefGoogle Scholar
  4. 4.
    Bannister AJ, Kouzarides T (2011) Regulation of chromatin by histone modifications. Cell Res 21(3):381–395CrossRefGoogle Scholar
  5. 5.
    Svinkina T et al (2015) Deep, quantitative coverage of the lysine acetylome using novel anti-acetyl-lysine antibodies and an optimized proteomic workflow. Mol Cell Proteomics 14(9):2429–2440CrossRefGoogle Scholar
  6. 6.
    Weinert BT et al (2018) Time-resolved analysis reveals rapid dynamics and broad scope of the CBP/p300 acetylome. Cell 174(1):231–244, e12CrossRefGoogle Scholar
  7. 7.
    Hornbeck PV et al (2015) PhosphoSitePlus, 2014: mutations, PTMs and recalibrations. Nucleic Acids Res 43(Database issue):D512–D520CrossRefGoogle Scholar
  8. 8.
    Gil J, Ramirez-Torres A, Encarnacion-Guevara S (2017) Lysine acetylation and cancer: a proteomics perspective. J Proteome 150:297–309CrossRefGoogle Scholar
  9. 9.
    Jones PA, Issa JP, Baylin S (2016) Targeting the cancer epigenome for therapy. Nat Rev Genet 17(10):630–641CrossRefGoogle Scholar
  10. 10.
    Lasko LM et al (2017) Discovery of a selective catalytic p300/CBP inhibitor that targets lineage-specific tumours. Nature 550(7674):128–132CrossRefGoogle Scholar
  11. 11.
    Fujisawa T, Filippakopoulos P (2017) Functions of bromodomain-containing proteins and their roles in homeostasis and cancer. Nat Rev Mol Cell Biol 18(4):246–262CrossRefGoogle Scholar
  12. 12.
    Dalvai M et al (2015) A scalable genome-editing-based approach for mapping multiprotein complexes in human cells. Cell Rep 13(3):621–633CrossRefGoogle Scholar
  13. 13.
    Doyon Y, Cote J (2016) Preparation and analysis of native chromatin-modifying complexes. Methods Enzymol 573:303–318CrossRefGoogle Scholar
  14. 14.
    Hockemeyer D et al (2009) Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases. Nat Biotechnol 27(9):851–857CrossRefGoogle Scholar
  15. 15.
    Neumann H et al (2009) A method for genetically installing site-specific acetylation in recombinant histones defines the effects of H3 K56 acetylation. Mol Cell 36(1):153–163CrossRefGoogle Scholar
  16. 16.
    Neumann H, Peak-Chew SY, Chin JW (2008) Genetically encoding N(epsilon)-acetyllysine in recombinant proteins. Nat Chem Biol 4(4):232–234CrossRefGoogle Scholar
  17. 17.
    Billon P et al (2017) Acetylation of PCNA sliding surface by Eco1 promotes genome stability through homologous recombination. Mol Cell 65(1):78–90CrossRefGoogle Scholar
  18. 18.
    Lin YY et al (2009) Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis. Cell 136(6):1073–1084CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.St. Patrick Research Group in Basic OncologyQuebec CityCanada
  2. 2.Laval University Cancer Research CenterQuebec CityCanada
  3. 3.Centre de Recherche du Centre Hospitalier Universitaire (CHU) de Québec-Université LavalQuebec CityCanada
  4. 4.Département de Médecine MoléculaireUniversité LavalQuebec CityCanada

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