Tackling aspecific side reactions during histone propionylation: The promise of reversing overpropionylation
| Autor: | Laura De Clerck, Paulien Meert, Elisabeth Govaert, Sander Willems, Maarten Dhaenens, Sebastiaan Dierickx, Dieter Deforce |
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| Rok vydání: | 2016 |
| Předmět: |
Propionylation
Proteomics 0301 basic medicine Technology PROTEINS Method optimization DNA repair Technical Brief Hydroxylamine Biology Arginine Peptide Mapping Biochemistry Anhydrides Histones Serine 03 medical and health sciences Humans Histone code Trypsin Amino Acid Sequence Epigenetics Molecular Biology Regulation of gene expression Mass spectrometry Lysine PEPTIDES MASS-SPECTROMETRY Hydrogen-Ion Concentration QUANTITATIVE PROTEOMIC ANALYSIS Peptide Fragments Chromatin Histone Code Chemistry Histone 030104 developmental biology Ammonium Hydroxide Solvents biology.protein Propionates Artifacts Protein Processing Post-Translational |
| Zdroj: | Proteomics PROTEOMICS |
| ISSN: | 1615-9853 |
| DOI: | 10.1002/pmic.201600045 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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