Protein ADP-ribosylation is a covalent, reversible posttranslational modification (PTM) catalyzed by ADP-ribosyltransferases (ARTs). Proteins can be either mono- or poly-ADP-ribosylated under a variety of physiological and pathological conditions. To understand the functional contribution of protein ADP-ribosylation to normal and disease/stress states, modified protein and corresponding ADP-ribose acceptor site identification is crucial. Since ADP-ribosylation is a transient and relatively low abundant PTM, systematic and accurate identification of ADP-ribose acceptor sites has only recently become feasible. This is due to the development of specific ADP-ribosylated protein/peptide enrichment methodologies, as well as technical advances in high-accuracy liquid chromatography–tandem mass spectrometry (LC-MS/MS). The standardized protocol described here allows the identification of ADP-ribose acceptor sites in in vitro ADP-ribosylated proteins and will, thus, contribute to the functional characterization of this important PTM.
CITATION STYLE
Leutert, M., Bilan, V., Gehrig, P., & Hottiger, M. O. (2017). Identification of ADP-ribose acceptor sites on in vitro modified proteins by liquid chromatography–tandem mass spectrometry. In Methods in Molecular Biology (Vol. 1608, pp. 137–148). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6993-7_10
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