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
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Kouzarides T (2000) Acetylation: a regulatory modification to rival phosphorylation? EMBO J 19(6):1176–1179
Allfrey VG, Mirsky AE (1964) Structural modifications of histones and their possible role in the regulation of RNA synthesis. Science 144(3618):559
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
Bannister AJ, Kouzarides T (2011) Regulation of chromatin by histone modifications. Cell Res 21(3):381–395
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
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
Hornbeck PV et al (2015) PhosphoSitePlus, 2014: mutations, PTMs and recalibrations. Nucleic Acids Res 43(Database issue):D512–D520
Gil J, Ramirez-Torres A, Encarnacion-Guevara S (2017) Lysine acetylation and cancer: a proteomics perspective. J Proteome 150:297–309
Jones PA, Issa JP, Baylin S (2016) Targeting the cancer epigenome for therapy. Nat Rev Genet 17(10):630–641
Lasko LM et al (2017) Discovery of a selective catalytic p300/CBP inhibitor that targets lineage-specific tumours. Nature 550(7674):128–132
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
Dalvai M et al (2015) A scalable genome-editing-based approach for mapping multiprotein complexes in human cells. Cell Rep 13(3):621–633
Doyon Y, Cote J (2016) Preparation and analysis of native chromatin-modifying complexes. Methods Enzymol 573:303–318
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
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
Neumann H, Peak-Chew SY, Chin JW (2008) Genetically encoding N(epsilon)-acetyllysine in recombinant proteins. Nat Chem Biol 4(4):232–234
Billon P et al (2017) Acetylation of PCNA sliding surface by Eco1 promotes genome stability through homologous recombination. Mol Cell 65(1):78–90
Lin YY et al (2009) Protein acetylation microarray reveals that NuA4 controls key metabolic target regulating gluconeogenesis. Cell 136(6):1073–1084
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
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
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