Quantitative assessment of chemical artefacts produced by propionylation of histones prior to mass spectrometry analysis

Research output: Contribution to journalComment/debate

Abstract

Histone PTMs play a crucial role in regulating chromatin structure and function, with impact on gene expression. MS is nowadays widely applied to study histone PTMs systematically. Because histones are rich in arginine and lysine, classical shot-gun approaches based on trypsin digestion are typically not employed for histone modifications mapping. Instead, different protocols of chemical derivatization of lysines in combination with trypsin have been implemented to obtain “Arg-C like” digestion products that are more suitable for LC-MS/MS analysis. Although widespread, these strategies have been recently described to cause various side reactions that result in chemical modifications prone to be misinterpreted as native histone marks. These artefacts can also interfere with the quantification process, causing errors in histone PTMs profiling. The work of Paternoster V. et al. is a quantitative assessment of methyl-esterification and other side reactions occurring on histones after chemical derivatization of lysines with propionic anhydride [Proteomics 2016, 16, 2059–2063]. The authors estimate the effect of different solvents, incubation times, and pH on the extent of these side reactions. The results collected indicate that the replacement of methanol with isopropanol or ACN not only blocks methyl-esterification, but also significantly reduces other undesired unspecific reactions. Carefully titrating the pH after propionic anhydride addition is another way to keep methyl-esterification under control. Overall, the authors describe a set of experimental conditions that allow reducing the generation of various artefacts during histone propionylation.

Original languageEnglish
Pages (from-to)1952-1954
Number of pages3
JournalProteomics
Volume16
Issue number14
DOIs
Publication statusPublished - Jul 1 2016

Fingerprint

Histones
Artifacts
Mass spectrometry
Mass Spectrometry
Esterification
Pulse time modulation
Histone Code
Lysine
Trypsin
Digestion
2-Propanol
Firearms
Proteomics
Chromatin
Methanol
Arginine
Chemical modification
Gene expression
Gene Expression
propionic anhydride

Keywords

  • Histone post-translational modifications
  • Lysine alkylation
  • Mass spectrometry
  • Methyl-esterification
  • Propionylation

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

Cite this

Quantitative assessment of chemical artefacts produced by propionylation of histones prior to mass spectrometry analysis. / Soldi, Monica; Cuomo, Alessandro; Bonaldi, Tiziana.

In: Proteomics, Vol. 16, No. 14, 01.07.2016, p. 1952-1954.

Research output: Contribution to journalComment/debate

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