Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Correction: Requirement for Serine-384 in Caspase-2 processing and activity.

Cell death & disease·2025
Same author

Genes That Associated with Action of ACTH-like Peptides with Neuroprotective Potential in Rat Brain Regions with Different Degrees of Ischemic Damage.

International journal of molecular sciences·2025
Same author

Inhibitors of Transketolase from <i>Mycobacterium tuberculosis</i> Targeted towards both the Diphosphate Binding Site and an Adjacent Hydrophobic Subsite.

Biochemistry. Biokhimiia·2025
Same author

ACTH-like Peptides Compensate Rat Brain Gene Expression Profile Disrupted by Ischemia a Day After Experimental Stroke.

Biomedicines·2025
Same author

Mechanism of PARP1 Elongation Reaction Revealed by Molecular Modeling.

Biochemistry. Biokhimiia·2024
Same author

Melanocortin Derivatives Induced Vascularization and Neuroglial Proliferation in the Rat Brain under Conditions of Cerebral Ischemia.

Current issues in molecular biology·2024

Related Experiment Video

Updated: Jun 10, 2025

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion
08:24

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion

Published on: July 27, 2021

1.4K

A New Approach for Studying Poly(ADP-Ribose) Polymerase Inhibitors Using Permeabilized Adherent Cells.

Stanislav I Shram1, Tatyana A Shcherbakova2, Tatyana V Abramova3

  • 1National Research Centre "Kurchatov Institute", Moscow, 123182, Russia. shram.img@yandex.ru.

Biochemistry. Biokhimiia
|October 17, 2024
PubMed
Summary

Researchers developed a new cell-based method to study poly(ADP-ribose) polymerase (PARP) activity and inhibition. This technique enhances the study of PARP inhibitors for drug development and disease treatment.

Keywords:
7,8-dimethylguanine7-methylguanineadherent cellscardiomyoblastscell permeabilizationchemical synthesis of purinesdigitonininhibitormolecular dockingpoly(ADP-ribose) polymerase

More Related Videos

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes
11:01

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Published on: August 24, 2021

2.7K
Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
07:26

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

9.5K

Related Experiment Videos

Last Updated: Jun 10, 2025

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion
08:24

Visualization of Replisome Encounters with an Antigen Tagged Blocking Lesion

Published on: July 27, 2021

1.4K
Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes
11:01

Automated Imaging and Analysis for the Quantification of Fluorescently Labeled Macropinosomes

Published on: August 24, 2021

2.7K
Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
07:26

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

9.5K

Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • Poly(ADP-ribose) polymerase (PARP) inhibitors show therapeutic potential across various diseases.
  • Understanding PARP's cellular context is crucial for optimizing inhibitor effectiveness.
  • Existing methods may not fully capture PARP's regulatory mechanisms in vivo.

Purpose of the Study:

  • To develop a novel cell-based model for studying poly(ADP-ribosyl)ation (PARylation) reactions.
  • To maintain nuclear localization of PARP and control substrate/inhibitor concentrations within the cellular environment.
  • To facilitate the study of PARP activity regulation and inhibitor screening.

Main Methods:

  • Utilized digitonin-permeabilized adherent cells to preserve cellular architecture.
  • Pre-treated cells to stabilize PARP in a DNA-bound state, preventing premature PARylation.
  • Quantified PARP activity by measuring poly(ADP-ribose) product via immunofluorescence in rat H9c2 cardiomyoblasts.

Main Results:

  • Successfully adapted digitonin-permeabilization for studying PARylation in a cellular context.
  • Validated the method using 3-aminobenzamide and novel 7-methylguanine derivatives.
  • Identified 7,8-dimethylguanine as a more potent PARP inhibitor than 7-methylguanine due to enhanced hydrophobic interactions.

Conclusions:

  • The proposed method provides a robust platform for investigating PARP activity regulation within a cellular environment.
  • This approach is suitable for high-throughput screening of potential PARP inhibitors.
  • Opens new avenues for understanding PARP function and developing targeted therapies.