Combined Metabolic and Chemical (CoMetChem) Labeling Using Stable Isotopesa Strategy to Reveal Site-Specific Histone Acetylation and Deacetylation Rates by LC–MS
datasetposted on 2021-09-14, 16:04 authored by Alienke van Pijkeren, Jörn Dietze, Alejandro Sánchez Brotons, Anna-Sophia Egger, Tim Lijster, Andrei Barcaru, Madlen Hotze, Philipp Kobler, Frank J. Dekker, Peter Horvatovich, Barbro N. Melgert, Mathias Ziegler, Kathrin Thedieck, Ines Heiland, Rainer Bischoff, Marcel Kwiatkowski
Histone acetylation is an important, reversible post-translational protein modification and a hallmark of epigenetic regulation. However, little is known about the dynamics of this process, due to the lack of analytical methods that can capture site-specific acetylation and deacetylation reactions. We present a new approach that combines metabolic and chemical labeling (CoMetChem) using uniformly 13C-labeled glucose and stable isotope-labeled acetic anhydride. Thereby, chemically equivalent, fully acetylated histone species are generated, enabling accurate relative quantification of site-specific lysine acetylation dynamics in tryptic peptides using high-resolution mass spectrometry. We show that CoMetChem enables site-specific quantification of the incorporation or loss of lysine acetylation over time, allowing the determination of reaction rates for acetylation and deacetylation. Thus, the CoMetChem methodology provides a comprehensive description of site-specific acetylation dynamics.
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using uniformly 13ctranslational protein modificationresolution mass spectrometrylabeled acetic anhydridecometchem methodology providesspecific histone acetylationcometchem enables sitespecific acetylation dynamicsspecific acetylationspecific quantificationlabeled glucoselysine acetylationstable isotopereversible postreveal sitereaction ratesnew approachepigenetic regulationcomprehensive descriptioncombines metaboliccombined metabolicchemically equivalentcapture siteanalytical methods