Profiling post-translational modifications of histones in human monocyte-derived macrophages

Pawel Olszowy, Maire Rose Donnelly, Chanho Lee, Pawel S Ciborowski

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: Histones and their post-translational modifications impact cellular function by acting as key regulators in the maintenance and remodeling of chromatin, thus affecting transcription regulation either positively (activation) or negatively (repression). In this study we describe a comprehensive, bottom-up proteomics approach to profiling post-translational modifications (acetylation, mono-, di- and tri-methylation, phosphorylation, biotinylation, ubiquitination, citrullination and ADP-ribosylation) in human macrophages, which are primary cells of the innate immune system. As our knowledge expands, it becomes more evident that macrophages are a heterogeneous population with potentially subtle differences in their responses to various stimuli driven by highly complex epigenetic regulatory mechanisms. Methods: To profile post-translational modifications (PTMs) of histones in macrophages we used two platforms of liquid chromatography and mass spectrometry. One platform was based on Sciex5600 TripleTof and the second one was based on VelosPro Orbitrap Elite ETD mass spectrometers. Results: We provide side-by-side comparison of profiling using two mass spectrometric platforms, ion trap and qTOF, coupled with the application of collisional induced and electron transfer dissociation. We show for the first time methylation of a His residue in macrophages and demonstrate differences in histone PTMs between those currently reported for macrophage cell lines and what we identified in primary cells. We have found a relatively low level of histone PTMs in differentiated but resting human primary monocyte derived macrophages. Conclusions: This study is the first comprehensive profiling of histone PTMs in primary human MDM. Our study implies that epigenetic regulatory mechanisms operative in transformed cell lines and primary cells are overlapping to a limited extent. Our mass spectrometric approach provides groundwork for the investigation of how histone PTMs contribute to epigenetic regulation in primary human macrophages.

Original languageEnglish (US)
Article number24
JournalProteome Science
Volume13
Issue number1
DOIs
StatePublished - Sep 24 2015

Fingerprint

Macrophages
Post Translational Protein Processing
Histones
Methylation
Epigenomics
Cells
Biotinylation
Acetylation
Transformed Cell Line
Phosphorylation
Chromatin Assembly and Disassembly
Immune system
Ubiquitination
Liquid chromatography
Mass spectrometers
Transcription
Liquid Chromatography
Proteomics
Adenosine Diphosphate
Chromatin

Keywords

  • Histones
  • Innate immunity
  • Macrophage
  • Mass spectrometry
  • Post-translational modification
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Profiling post-translational modifications of histones in human monocyte-derived macrophages. / Olszowy, Pawel; Donnelly, Maire Rose; Lee, Chanho; Ciborowski, Pawel S.

In: Proteome Science, Vol. 13, No. 1, 24, 24.09.2015.

Research output: Contribution to journalArticle

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