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Title:
Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry | SpringerLink
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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...
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ScholarlyArticle:
headline:Quantitation of Poly(ADP-Ribose) by Isotope Dilution Mass Spectrometry
pageEnd:18
pageStart:3
image:https://media.springernature.com/w153/springer-static/cover/book/978-1-4939-6993-7.jpg
genre:
Springer Protocols
isPartOf:
name:Poly(ADP-Ribose) Polymerase
isbn:
978-1-4939-6993-7
978-1-4939-6992-0
type:Book
publisher:
name:Springer New York
logo:
url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
type:ImageObject
type:Organization
author:
name:Tabea Zubel
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
type:Person
name:Rita Martello
affiliation:
name:University of Copenhagen
address:
name:Department of Proteomics, Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark
type:PostalAddress
type:Organization
type:Person
name:Alexander Bürkle
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
type:Person
name:Aswin Mangerich
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
email:[email protected]
type:Person
keywords:Poly(ADP-ribose), PARP, ARTD, Mass spectrometry, UPLC, HPLC
description: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.
datePublished:2017
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Book:
name:Poly(ADP-Ribose) Polymerase
isbn:
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978-1-4939-6992-0
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name:Springer New York
logo:
url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
type:ImageObject
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
name:University of Copenhagen
address:
name:Department of Proteomics, Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark
type:PostalAddress
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
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Person:
name:Tabea Zubel
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
name:Rita Martello
affiliation:
name:University of Copenhagen
address:
name:Department of Proteomics, Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark
type:PostalAddress
type:Organization
name:Alexander Bürkle
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
name:Aswin Mangerich
affiliation:
name:University of Konstanz
address:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
type:PostalAddress
type:Organization
email:[email protected]
PostalAddress:
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
name:Department of Proteomics, Faculty of Health and Medical Sciences, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen, Denmark
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
name:Molecular Toxicology Group, Department of Biology, University of Konstanz, Konstanz, Germany
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