Skip to main content

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

  • Protocol
  • First Online:
Protein Acetylation

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

Abstract

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.

Jean-Philippe Lambert and Jacques Côté are both corresponding author.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Kouzarides T (2000) Acetylation: a regulatory modification to rival phosphorylation? EMBO J 19(6):1176–1179

    Article  CAS  Google Scholar 

  2. Allfrey VG, Mirsky AE (1964) Structural modifications of histones and their possible role in the regulation of RNA synthesis. Science 144(3618):559

    Article  CAS  Google Scholar 

  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):805–812

    Article  CAS  Google Scholar 

  4. Bannister AJ, Kouzarides T (2011) Regulation of chromatin by histone modifications. Cell Res 21(3):381–395

    Article  CAS  Google Scholar 

  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):2429–2440

    Article  CAS  Google Scholar 

  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, e12

    Article  CAS  Google Scholar 

  7. Hornbeck PV et al (2015) PhosphoSitePlus, 2014: mutations, PTMs and recalibrations. Nucleic Acids Res 43(Database issue):D512–D520

    Article  CAS  Google Scholar 

  8. Gil J, Ramirez-Torres A, Encarnacion-Guevara S (2017) Lysine acetylation and cancer: a proteomics perspective. J Proteome 150:297–309

    Article  CAS  Google Scholar 

  9. Jones PA, Issa JP, Baylin S (2016) Targeting the cancer epigenome for therapy. Nat Rev Genet 17(10):630–641

    Article  CAS  Google Scholar 

  10. Lasko LM et al (2017) Discovery of a selective catalytic p300/CBP inhibitor that targets lineage-specific tumours. Nature 550(7674):128–132

    Article  CAS  Google Scholar 

  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):246–262

    Article  CAS  Google Scholar 

  12. Dalvai M et al (2015) A scalable genome-editing-based approach for mapping multiprotein complexes in human cells. Cell Rep 13(3):621–633

    Article  CAS  Google Scholar 

  13. Doyon Y, Cote J (2016) Preparation and analysis of native chromatin-modifying complexes. Methods Enzymol 573:303–318

    Article  CAS  Google Scholar 

  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):851–857

    Article  CAS  Google Scholar 

  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):153–163

    Article  CAS  Google Scholar 

  16. Neumann H, Peak-Chew SY, Chin JW (2008) Genetically encoding N(epsilon)-acetyllysine in recombinant proteins. Nat Chem Biol 4(4):232–234

    Article  CAS  Google Scholar 

  17. Billon P et al (2017) Acetylation of PCNA sliding surface by Eco1 promotes genome stability through homologous recombination. Mol Cell 65(1):78–90

    Article  CAS  Google Scholar 

  18. Lin YY et al (2009) Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis. Cell 136(6):1073–1084

    Article  CAS  Google Scholar 

Download references

Acknowledgments

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.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Jean-Philippe Lambert or Jacques Côté .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

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

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lashgari, A., Lambert, JP., Côté, J. (2019). Measurement and Analysis of Lysine Acetylation by KAT Complexes In Vitro and In Vivo. In: Brosh, Jr., R. (eds) Protein Acetylation. Methods in Molecular Biology, vol 1983. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9434-2_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9434-2_5

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9433-5

  • Online ISBN: 978-1-4939-9434-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics