A Proteomic Approach to Analyze the Aspirin-mediated Lysine Acetylome

Michael H Tatham, Christian Cole, Paul Scullion, Ross Wilkie, Nicholas J. Westwood, Lesley A. Stark, Ronald T Hay (Lead / Corresponding author)

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Abstract

Aspirin, or acetylsalicylic acid is widely used to control pain, inflammation and fever. Important to this function is its ability to irreversibly acetylate cyclooxygenases at active site serines. Aspirin has the potential to acetylate other amino-acid side-chains, leading to the possibility that aspirin-mediated lysine acetylation could explain some of its as-yet unexplained drug actions or side-effects. Using isotopically labeled aspirin-d3, in combination with acetylated lysine purification and LC-MS/MS, we identified over 12000 sites of lysine acetylation from cultured human cells. Although aspirin amplifies endogenous acetylation signals at the majority of detectable endogenous sites, cells tolerate aspirin mediated acetylation very well unless cellular deacetylases are inhibited. Although most endogenous acetylations are amplified by orders of magnitude, lysine acetylation site occupancies remain very low even after high doses of aspirin. This work shows that while aspirin has enormous potential to alter protein function, in the majority of cases aspirin-mediated acetylations do not accumulate to levels likely to elicit biological effects. These findings are consistent with an emerging model for cellular acetylation whereby stoichiometry correlates with biological relevance, and deacetylases act to minimize the biological consequences non-specific chemical acetylations.
Original languageEnglish
Pages (from-to)310-326
Number of pages17
JournalMolecular & Cellular Proteomics
Volume16
Issue number2
Early online date2 Dec 2016
DOIs
Publication statusPublished - 1 Feb 2017

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Keywords

  • Acetylation
  • Aspirin
  • Binding Sites
  • Chromatography, Liquid
  • HeLa Cells
  • Histone Deacetylases
  • Humans
  • Isotope Labeling
  • Lysine
  • Proteome
  • Proteomics
  • Tandem Mass Spectrometry

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