Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry

Zubel, Tabea; Martello, Rita; Bürkle, Alexander; Mangerich, Aswin

Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry

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Datum

2017

Publikationstyp

Beitrag zu einem Sammelband

Publikationsstatus

Published

Erschienen in

TULIN, Alexei V., ed.. Poly(ADP-Ribose) Polymerase : methods and protocols. second edition. New York, NY: Humana Press, 2017, pp. 3-18. Methods in Molecular Biology. 1608. ISBN 978-1-4939-6992-0. Available under: doi: 10.1007/978-1-4939-6993-7_1

Zusammenfassung

Poly(ADP-ribosyl)ation (PARylation), i.e., the formation of the nucleic acid-like biopolymer poly(ADP-ribose) (PAR), is an essential posttranslational modification carried out by poly(ADP-ribose) polymerases (PARPs). While PAR levels are low under physiological conditions, they can transiently increase more than 100-fold upon induction of genotoxic stress. The accurate quantitation of cellular PAR with high sensitivity is of critical importance to understand the role of PARylation in cellular physiology and pathophysiology and to determine the pharmacodynamic efficiencies of clinically relevant PARP inhibitors, which represent a novel class of promising chemotherapeutics. Previously, we have developed a bioanalytical platform based on isotope dilution mass spectrometry (LC-MS/MS) to quantify cellular PAR with unequivocal chemical specificity in absolute terms with femtomol sensitivity (Martello et al. ACS Chem Biol 8(7):1567-1575, 2013). This method enables the analysis of steady-state levels, as well as stress-induced levels of PAR in various biological systems including cell lines, mouse tissues, and primary human lymphocytes. It has a wide range of potential applications in basic research, as well as in drug development (Martello et al. ACS Chem Biol 8(7):1567-1575, 2013; Mangerich et al. Toxicol Lett 244:56-71, 2016). Here, we present an improved and adjusted version of the original protocol by Martello/Mangerich et al., which uses UPLC-MS/MS instrumentation.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

ZUBEL, Tabea, Rita MARTELLO, Alexander BÜRKLE, Aswin MANGERICH, 2017. Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry. In: TULIN, Alexei V., ed.. Poly(ADP-Ribose) Polymerase : methods and protocols. second edition. New York, NY: Humana Press, 2017, pp. 3-18. Methods in Molecular Biology. 1608. ISBN 978-1-4939-6992-0. Available under: doi: 10.1007/978-1-4939-6993-7_1

BibTex

@incollection{Zubel2017Quant-40288,
  year={2017},
  doi={10.1007/978-1-4939-6993-7_1},
  title={Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry},
  edition={second edition},
  number={1608},
  isbn={978-1-4939-6992-0},
  publisher={Humana Press},
  address={New York, NY},
  series={Methods in Molecular Biology},
  booktitle={Poly(ADP-Ribose) Polymerase : methods and protocols},
  pages={3--18},
  editor={Tulin, Alexei V.},
  author={Zubel, Tabea and Martello, Rita and Bürkle, Alexander and Mangerich, Aswin}
}

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    <dcterms:abstract xml:lang="eng">Poly(ADP-ribosyl)ation (PARylation), i.e., the formation of the nucleic acid-like biopolymer poly(ADP-ribose) (PAR), is an essential posttranslational modification carried out by poly(ADP-ribose) polymerases (PARPs). While PAR levels are low under physiological conditions, they can transiently increase more than 100-fold upon induction of genotoxic stress. The accurate quantitation of cellular PAR with high sensitivity is of critical importance to understand the role of PARylation in cellular physiology and pathophysiology and to determine the pharmacodynamic efficiencies of clinically relevant PARP inhibitors, which represent a novel class of promising chemotherapeutics. Previously, we have developed a bioanalytical platform based on isotope dilution mass spectrometry (LC-MS/MS) to quantify cellular PAR with unequivocal chemical specificity in absolute terms with femtomol sensitivity (Martello et al. ACS Chem Biol 8(7):1567-1575, 2013). This method enables the analysis of steady-state levels, as well as stress-induced levels of PAR in various biological systems including cell lines, mouse tissues, and primary human lymphocytes. It has a wide range of potential applications in basic research, as well as in drug development (Martello et al. ACS Chem Biol 8(7):1567-1575, 2013; Mangerich et al. Toxicol Lett 244:56-71, 2016). Here, we present an improved and adjusted version of the original protocol by Martello/Mangerich et al., which uses UPLC-MS/MS instrumentation.</dcterms:abstract>
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