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Detecting and Quantifying pADPr In Vivo.

Yi-Chen Lai1,2, Rajesh K Aneja3,4, Margaret A Satchell4

  • 1Department of Pediatrics, Baylor College of Medicine, Houston, TX, USA. ylai@bcm.edu.

Methods in Molecular Biology (Clifton, N.J.)
|July 12, 2017
PubMed
Summary
This summary is machine-generated.

Poly(ADP-ribose) polymerases (PARP) play roles in cell health and injury. Detecting poly-ADP-ribosylation (pADPr) can offer insights into diseases and guide PARP inhibitor therapies.

Keywords:
ADP-ribose polymerasePoly(ADP-ribose) polymerasePoly(ADP-ribose) polymersPoly(ADP-ribose) synthetasePoly-ADP-ribosylation

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Poly(ADP-ribose) polymerases (PARP) are crucial enzymes involved in DNA repair and cellular homeostasis.
  • PARP overactivation, indicated by increased poly-ADP-ribosylation (pADPr), contributes to NAD+ depletion and mitochondrial dysfunction, potentially leading to cell death.
  • Elevated pADPr levels are implicated in the pathology of various diseases.

Purpose of the Study:

  • To review non-isotopic immunodetection methods for quantifying pADPr.
  • To highlight the significance of monitoring PARP-1 activation via pADPr detection for understanding disease mechanisms and therapeutic drug monitoring.
  • To discuss the utility of pADPr quantification in the context of PARP inhibitor efficacy.

Main Methods:

  • Western blotting to detect poly-ADP-ribosylated proteins.
  • Immunohistochemistry for cellular localization of pADPr.
  • Enzyme-linked immunoassay (ELISA) for quantifying pADPr.
  • Small-scale two-dimensional gel electrophoresis for pADPr analysis.

Main Results:

  • Several non-isotopic immunodetection methods are available for quantifying pADPr.
  • These methods allow for the assessment of PARP activity and its consequences.
  • The discussed techniques provide options for both qualitative and quantitative analysis of pADPr.

Conclusions:

  • Monitoring PARP activation through pADPr quantification is valuable for disease research.
  • pADPr detection aids in understanding the role of PARP in cellular dysfunction.
  • These methods can facilitate the monitoring of PARP inhibitor therapies in clinical settings.