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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