title: Tackling aspecific side reactions during histone propionylation: The promise of reversing overpropionylation.
abstract: Histone proteins are essential elements for DNA packaging. Moreover, the PTMs that are extremely abundant on these proteins, contribute in modeling chromatin structure and recruiting enzymes involved in gene regulation, DNA repair and chromosome condensation. This fundamental aspect, together with the epigenetic inheritance of histone PTMs, underlines the importance of having biochemical techniques for their characterization. Over the past two decades, significant improvements in mass accuracy and resolution of mass spectrometers have made LC-coupled MS the strategy of choice for accurate identification and quantification of protein PTMs. Nevertheless, in previous work we disclosed the limitations and biases of the most widely adopted sample preparation protocols for histone propionylation, required prior to bottom-up MS analysis. In this work, however, we put forward a new specific and efficient propionylation strategy by means of propionic anhydride. In this method, aspecific overpropionylation at serine (S), threonine (T) and tyrosine (Y) is reversed by adding hydroxylamine (HA). We recommend using this method for future analysis of histones through bottom-up MS.
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Paulien
authors.initials: P
authors.lastname: Meert
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Sebastiaan
authors.initials: S
authors.lastname: Dierickx
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Govaert
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Laura
authors.initials: L
authors.lastname: De Clerck
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Sander
authors.initials: S
authors.lastname: Willems
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: Dhaenens
authors.affiliation: Laboratory of Pharmaceutical Biotechnology, Ghent University, Ghent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
mesh_terms: D000595:Amino Acid Sequence; D064753:Ammonium Hydroxide / Q000737:chemistry; D000812:Anhydrides / Q000737:chemistry* / Q000378:metabolism; D001120:Arginine / Q000737:chemistry / Q000378:metabolism; D016477:Artifacts; D042421:Histone Code; D006657:Histones / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019811:Hydroxylamine / Q000737:chemistry; D008239:Lysine / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000592:standards; D010446:Peptide Fragments / Q000032:analysis*; D010449:Peptide Mapping; D011422:Propionates / Q000737:chemistry* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D012997:Solvents / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
chemical_list: D000812:Anhydrides; D006657:Histones; D010446:Peptide Fragments; D011422:Propionates; D012997:Solvents; D019811:Hydroxylamine; D064753:Ammonium Hydroxide; D001120:Arginine; C096126:propionic anhydride; D014357:Trypsin; D008239:Lysine
keywords: Histone; Mass spectrometry; Method optimization; Propionylation; Technology