Elsevier

Clinical Mass Spectrometry

Volume 15, January 2020, Pages 54-60
Clinical Mass Spectrometry

Determining histone H4 acetylation patterns in human peripheral blood mononuclear cells using mass spectrometry

https://doi.org/10.1016/j.clinms.2019.08.001Get rights and content
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Highlights

  • Histone acetylations can be precisely quantified in hPBMCs.

  • Histone H4 acetylation changes specifically during ageing.

  • Age-related changes are evolutionary conserved.

Abstract

Misregulated chromatin remodeling via erroneous histone acetylation has been linked with multiple diseases including many age-associated maladies. Previous studies on histone acetylation have utilized specific histone acetylation antibodies as a primary quantification tool. However, antibodies that target specific histone modifications frequently lack sufficient sensitivity and specificity, making signal quantification and data interpretation difficult. Here, we used mass spectrometry to quantify the acetylation patterns of histone H4 in peripheral blood mononuclear cells (PBMCs) from human patients of varying age. We observed that mono-acetylation of H4K16ac is lower in midlife human PBMCs, and that various changes occur in the H4 acetylation signature using PBMCs prepared from old patients. Our data corroborates previous work suggesting that ageing might be characterized by specific changes in the acetylation states of histone H4 and shows that targeted mass spectrometry together with appropriate quantification methods can be used to sensitively measure such changes.

Abbreviations

Ac
Acetylation
Am
Ante meridiem
DTT
1,4-Dithiotreitol
EDTA
Ethylenediaminetetraacetic acid
H2SO4
Sulfuric acid
H4
Histone 4
H4K5 (as example of all modifications)
Lysine 5 of histone 4
HEPES
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
hPBMC
Human peripheral blood mononuclear cells
K5, K8, K12, K16
Lysine 5, Lysine 8, Lysine 12, Lysine 16
KAT
Lysine acetyltransferase
KCl
Potassium chloride
KDAC
Lysine deacetylases
KDACi
Lysine deacetylase inhibitors
M
Molar
MgCl2
Magnesium chloride
mPBMC
Mouse peripheral blood mononuclear cell
NaCl
Sodium chloride
NH4HCO3/Ambic
Ammonium bicarbonate
PBMC
Peripheral blood mononuclear cell
PBS
Phosphate-buffered saline
pH
Potential of hydrogen
TCA
Trichloroacetic acid
TFA
Trifluoroacetic acid

Cited by (0)

1

Co-senior authors.

2

Present address: Klinik für Pneumologie, University of Zürich, Germany.