Publikation:

Real-time monitoring of PARP1-dependent PARylation by ATR-FTIR spectroscopy

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2020

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http://nbn-resolving.de/urn:nbn:de:bsz:352-2-8wux0ubfp63x8
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https://doi.org/10.1038/s41467-020-15858-w
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CC BY 4.0

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Biologie: Publikationen
Chemie: Publikationen
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Published

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Nature Communications. Nature Publishing Group. 2020, 11(1), 2174. eISSN 2041-1723. Available under: doi: 10.1038/s41467-020-15858-w

Zusammenfassung

Poly-ADP-ribosylation (PARylation) is a fully reversible post-translational modification with key roles in cellular physiology. Due to the multi-domain structure of poly(ADP-ribose) polymerase-1 (PARP1) and the highly dynamic nature of the PARylation reaction, studies on the biochemical mechanism and structural dynamics remain challenging. Here, we report label-free, time-resolved monitoring of PARP1-dependent PARylation using ATR-FTIR spectroscopy. This includes PARP1 activation by binding to DNA strand break models, NAD+ substrate binding, PAR formation, and dissociation of automodified PARP1 from DNA. Analyses of PARP1 activation at different DNA models demonstrate a strong positive correlation of PARylation and PARP1 dissociation, with the strongest effects observed for DNA nicks and 3’ phosphorylated ends. Moreover, by examining dynamic structural changes of PARP1, we reveal changes in the secondary structure of PARP1 induced by NAD+ and PARP inhibitor binding. In summary, this approach enables holistic and dynamic insights into PARP1-dependent PARylation with molecular and temporal resolution.

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570 Biowissenschaften, Biologie

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ISO 690KRÜGER, Annika, Alexander BÜRKLE, Karin HAUSER, Aswin MANGERICH, 2020. Real-time monitoring of PARP1-dependent PARylation by ATR-FTIR spectroscopy. In: Nature Communications. Nature Publishing Group. 2020, 11(1), 2174. eISSN 2041-1723. Available under: doi: 10.1038/s41467-020-15858-w
BibTex
@article{Kruger2020-05-01Realt-49384,
  year={2020},
  doi={10.1038/s41467-020-15858-w},
  title={Real-time monitoring of PARP1-dependent PARylation by ATR-FTIR spectroscopy},
  number={1},
  volume={11},
  journal={Nature Communications},
  author={Krüger, Annika and Bürkle, Alexander and Hauser, Karin and Mangerich, Aswin},
  note={Article Number: 2174}
}
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