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

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.5K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.5K

You might also read

Related Articles

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

Sort by
Same author

Deciphering cytokine-driven ADP-ribosylation signaling networks via Af1521-based mass spectrometry analysis of labile Glu/Asp-linkages.

Nature communications·2026
Same author

Structure of a contractile injection system in Salmonella enterica subsp. salamae.

Nature communications·2026
Same author

UFMylation of Pyruvate Dehydrogenase Regulates Mitochondrial Metabolism.

bioRxiv : the preprint server for biology·2026
Same author

Higher-Throughput Proteome Profiling Enabled by Parallelized Pre-Accumulation and Optimized Ion Processing in the Orbitrap Astral Zoom Mass Spectrometer.

Molecular & cellular proteomics : MCP·2026
Same author

PARG regulates the proteasomal degradation of TARG1.

Cell reports·2026
Same author

PARP1 auto-modification promotes faithful Okazaki fragment processing and limits replication fork speed.

Molecular cell·2025

Related Experiment Video

Updated: Dec 8, 2025

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time
08:33

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

Published on: March 11, 2021

2.2K

Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation.

Sara C Buch-Larsen1, Ivo A Hendriks1, Jean M Lodge2

  • 1Proteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.

Cell Reports
|September 23, 2020
PubMed
Summary

We introduce activated ion electron transfer dissociation (AI-ETD) for mass spectrometry to analyze ADP-ribosylation (ADPr) in cells. This method enhances the identification of ADPr sites, offering new insights into cellular processes.

Keywords:
ADP-ribosylationAI-ETDGolgi apparatusPARP14PARP8endoplasmic reticulumglycosylationmass spectrometryphysiological ADPrproteomics

More Related Videos

Chemical Triphosphorylation of Oligonucleotides
13:19

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

3.9K
Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
09:40

Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

Published on: September 20, 2011

18.3K

Related Experiment Videos

Last Updated: Dec 8, 2025

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time
08:33

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

Published on: March 11, 2021

2.2K
Chemical Triphosphorylation of Oligonucleotides
13:19

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

3.9K
Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
09:40

Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

Published on: September 20, 2011

18.3K

Area of Science:

  • Biochemistry
  • Proteomics
  • Molecular Biology

Background:

  • ADP-ribosylation (ADPr) is a crucial post-translational modification involved in numerous cellular functions.
  • Analyzing physiological ADPr using mass spectrometry (MS) is challenging due to technical limitations and the need for unbiased enrichment strategies.

Purpose of the Study:

  • To evaluate the efficacy of activated ion electron transfer dissociation (AI-ETD) coupled with Af1521 enrichment for MS-based analysis of physiological ADP-ribosylation.
  • To benchmark AI-ETD against existing methods like ETD and EThcD for identifying ADP-ribosylated peptides and sites.

Main Methods:

  • Utilized an unbiased Af1521 enrichment strategy for ADP-ribosylated peptides.
  • Employed activated ion electron transfer dissociation (AI-ETD) for mass spectrometry-based proteomics.
  • Compared AI-ETD performance with traditional ETD and EThcD methods.

Main Results:

  • AI-ETD identified 120% and 28% more ADPr peptides than ETD and EThcD, respectively, from cells under oxidative stress.
  • Over 5,000 unique ADPr sites were identified, including 450 sites on low-abundant proteins under physiological conditions.
  • Discovered in vivo cysteine modifications on PARP8 and tyrosine modifications on PARP14, suggesting specific enzymatic roles.

Conclusions:

  • AI-ETD is a powerful and efficient technique for MS-based analysis of physiological ADP-ribosylation.
  • This approach significantly advances the ability to profile ADPr modifications, even on low-abundance proteins.
  • The findings provide novel insights into the regulation and function of ADP-ribosylation in cellular processes.