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)

Research output: Contribution to journalArticle

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127 Downloads (Pure)

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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Acetylation
Proteomics
Aspirin
Lysine
acetylsalicylic acid lysinate
Aptitude
Prostaglandin-Endoperoxide Synthases
Serine
Stoichiometry
Cultured Cells
Catalytic Domain
Purification
Fever
Inflammation
Cells
Amino Acids
Pain

Keywords

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

Cite this

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title = "A Proteomic Approach to Analyze the Aspirin-mediated Lysine Acetylome",
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.",
keywords = "Acetylation, Aspirin, Binding Sites, Chromatography, Liquid, HeLa Cells, Histone Deacetylases, Humans, Isotope Labeling, Lysine, Proteome, Proteomics, Tandem Mass Spectrometry",
author = "Tatham, {Michael H} and Christian Cole and Paul Scullion and Ross Wilkie and Westwood, {Nicholas J.} and Stark, {Lesley A.} and Hay, {Ronald T}",
note = "This work is supported by Cancer Research UK Grant C434/A13067 (M.H.T & R.T.H) and Wellcome Trust Grant 098391/Z/12/7 (R.T.H.).",
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A Proteomic Approach to Analyze the Aspirin-mediated Lysine Acetylome. / Tatham, Michael H; Cole, Christian; Scullion, Paul; Wilkie, Ross; Westwood, Nicholas J.; Stark, Lesley A.; Hay, Ronald T (Lead / Corresponding author).

In: Molecular & Cellular Proteomics, Vol. 16, No. 2, 01.02.2017, p. 310-326.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A Proteomic Approach to Analyze the Aspirin-mediated Lysine Acetylome

AU - Tatham, Michael H

AU - Cole, Christian

AU - Scullion, Paul

AU - Wilkie, Ross

AU - Westwood, Nicholas J.

AU - Stark, Lesley A.

AU - Hay, Ronald T

N1 - This work is supported by Cancer Research UK Grant C434/A13067 (M.H.T & R.T.H) and Wellcome Trust Grant 098391/Z/12/7 (R.T.H.).

PY - 2017/2/1

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N2 - 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.

AB - 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.

KW - Acetylation

KW - Aspirin

KW - Binding Sites

KW - Chromatography, Liquid

KW - HeLa Cells

KW - Histone Deacetylases

KW - Humans

KW - Isotope Labeling

KW - Lysine

KW - Proteome

KW - Proteomics

KW - Tandem Mass Spectrometry

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