TY - JOUR
T1 - Combined Metabolic and Chemical (CoMetChem) Labeling Using Stable Isotopes—a Strategy to Reveal Site-Specific Histone Acetylation and Deacetylation Rates by LC-MS
AU - van Pijkeren, Alienke
AU - Dietze, Jörn
AU - Brotons, Alejandro Sánchez
AU - Egger, Anna-Sophia
AU - Lijster, Tim
AU - Barcaru, Andrei
AU - Hotze, Madlen
AU - Kobler, Philipp
AU - Dekker, Frank J
AU - Horvatovich, Peter
AU - Melgert, Barbro N
AU - Ziegler, Mathias
AU - Thedieck, Kathrin
AU - Heiland, Ines
AU - Bischoff, Rainer
AU - Kwiatkowski, Marcel
N1 - Funding Information:
M.K. thanks the European Respiratory Society (ERS, RESPIRE3, project reference: R3201703-00121), the University of Innsbruck (project no: 316826), and the Tyrolian Research Fund (project no: 18903) for financial support. J.D., I.H., and M.Z. have been supported by the Norwegian Research Foundation (ES633272 and 302314). We acknowledge support from the MESI-STRAT project (grant agreement 754688 to K.T., I.H., and M.Z.) and the PoLiMeR Innovative Training Network (Marie Sklodowska-Curie grant agreement 812616 to K.T. and M.Z.), which received funding from the European Union Horizon 2020 Research and Innovation Program. A.S.B. and P.H. have received funding from the Netherlands X-omics Initiative, which is partially funded by the NWO (project no: 184.034.019). F.J.D. thanks the European Research Council for an ERC starting grant (309782) and Netherlands Organization of Scientific Research (NWO, VIDI grant: 723.012.005) for financial support. The LC–MS data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD027338.
Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society
PY - 2021/9/28
Y1 - 2021/9/28
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85115999817&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.1c01359
DO - 10.1021/acs.analchem.1c01359
M3 - Article
C2 - 34519498
SN - 0003-2700
VL - 93
SP - 12872
EP - 12880
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 38
ER -