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Profiling of the ADP-Ribosylome in Living Cells

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2022

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Angewandte Chemie International Edition. Wiley. 2022, 61(18), e202200977. ISSN 1433-7851. eISSN 1521-3773. Available under: doi: 10.1002/anie.202200977

Zusammenfassung

Post-translational modification (PTM) with ADP-ribose and poly(ADP-ribose) using nicotinamide adenine dinucleotide (NAD+ ) as substrate is involved in the regulation of numerous cellular pathways in eukaryotes, notably the response to DNA damage caused by cellular stress. Nevertheless, due to intrinsic properties of NAD+ e.g., high polarity and associated poor cell passage, these PTMs are difficult to characterize in cells. Here, two new NAD+ derivatives are presented, which carry either a fluorophore or an affinity tag and, in combination with developed methods for mild cell delivery, allow studies in living human cells. We show that this approach allows not only the imaging of ADP-ribosylation in living cells but also the proteome-wide analysis of cellular adaptation by protein ADP-ribosylation as a consequence of environmental changes such as H2 O2 -induced oxidative stress or the effect of the approved anti-cancer drug olaparib. Our results therefore pave the way for further functional and clinical studies of the ADP-ribosylated proteome in living cells in health and disease.

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ISO 690LEHNER, Maike, Sonja RIETH, Eva HÖLLMÜLLER, Daniel SPLIESGAR, Bastian MERTES, Florian STENGEL, Andreas MARX, 2022. Profiling of the ADP-Ribosylome in Living Cells. In: Angewandte Chemie International Edition. Wiley. 2022, 61(18), e202200977. ISSN 1433-7851. eISSN 1521-3773. Available under: doi: 10.1002/anie.202200977
BibTex
@article{Lehner2022-04-25Profi-57045,
  year={2022},
  doi={10.1002/anie.202200977},
  title={Profiling of the ADP-Ribosylome in Living Cells},
  number={18},
  volume={61},
  issn={1433-7851},
  journal={Angewandte Chemie International Edition},
  author={Lehner, Maike and Rieth, Sonja and Höllmüller, Eva and Spliesgar, Daniel and Mertes, Bastian and Stengel, Florian and Marx, Andreas},
  note={A.M. acknowledges support from the German Research Foundation (DFG grant MA 2288/21); F.S. is grateful for funding from the German Science Foundation Emmy Noether Programme (STE 2517/1-1) Article Number: e202200977}
}
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    <dcterms:abstract xml:lang="eng">Post-translational modification (PTM) with ADP-ribose and poly(ADP-ribose) using nicotinamide adenine dinucleotide (NAD&lt;sup&gt;+&lt;/sup&gt; ) as substrate is involved in the regulation of numerous cellular pathways in eukaryotes, notably the response to DNA damage caused by cellular stress. Nevertheless, due to intrinsic properties of NAD&lt;sup&gt;+&lt;/sup&gt; e.g., high polarity and associated poor cell passage, these PTMs are difficult to characterize in cells. Here, two new NAD+ derivatives are presented, which carry either a fluorophore or an affinity tag and, in combination with developed methods for mild cell delivery, allow studies in living human cells. We show that this approach allows not only the imaging of ADP-ribosylation in living cells but also the proteome-wide analysis of cellular adaptation by protein ADP-ribosylation as a consequence of environmental changes such as H&lt;sub&gt;2&lt;/sub&gt; O&lt;sub&gt;2&lt;/sub&gt; -induced oxidative stress or the effect of the approved anti-cancer drug olaparib. Our results therefore pave the way for further functional and clinical studies of the ADP-ribosylated proteome in living cells in health and disease.</dcterms:abstract>
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A.M. acknowledges support from the German Research Foundation (DFG grant MA 2288/21); F.S. is grateful for funding from the German Science Foundation Emmy Noether Programme (STE 2517/1-1)
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