Combined Metabolic and Chemical (CoMetChem) Labeling Using Stable Isotopes—a Strategy to Reveal Site-Specific Histone Acetylation and Deacetylation Rates by LC-MS

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

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Abstract

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.

Original languageEnglish
Pages (from-to)12872-12880
Number of pages9
JournalAnalytical Chemistry
Volume93
Issue number38
Early online date14-Sep-2021
DOIs
Publication statusPublished - 28-Sep-2021

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