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 Modifications in the RER01:26

Protein Modifications in the RER

5.6K
Modification of secretory and transmembrane proteins entering the rough ER begins in the ER lumen. These modifications aid in protein folding and stabilize the acquired tertiary structure. Protein modifications in the rough ER co-occur at different stages of protein folding.
Broadly, these modifications can be categorized into four main categories — glycosylation, formation of disulfide bonds, assembly of protein subunits, and specific proteolytic cleavages like removal of signal...
5.6K
Mass Spectrometry: Molecular Fragmentation Overview01:20

Mass Spectrometry: Molecular Fragmentation Overview

5.2K
The ionization of a molecule into a molecular ion inside the mass spectrometer causes instability in the molecule's structure due to the loss of an electron. This eventually leads to the fragmentation or breaking of some bonds in the molecule. The fragmentation occurs predominantly at specific bonds to yield relatively stable fragments.
One type of fragmentation pattern is the cleavage of a single bond in the molecular ion. The cleavage leads to a radical and a cation. The cleavage can occur at...
5.2K
Protein Folding01:22

Protein Folding

112.3K
Overview
112.3K
Protein Folding01:25

Protein Folding

8.8K
Proteins are chains of amino acids linked together by peptide bonds. Upon synthesis, a protein folds into a three-dimensional conformation, critical to its biological function. Interactions between its constituent amino acids guide protein folding, and hence the protein structure is primarily dependent on its amino acid sequence.
Protein Structure Is Critical to Its Biological Function
Proteins perform a wide range of biological functions such as catalyzing chemical reactions, providing...
8.8K
Preparation and Reactions of Thiols02:33

Preparation and Reactions of Thiols

6.7K
Thiols are prepared using the hydrosulfide anion as a nucleophile in a nucleophilic substitution reaction with alkyl halides. For instance, bromobutane reacts with sodium hydrosulfide to give butanethiol.
6.7K
Protein and Protein Structure02:15

Protein and Protein Structure

71.5K
Proteins are one of the most abundant organic molecules in living systems and have the most diverse range of functions of all macromolecules. Proteins may be structural, regulatory, contractile, or protective. They may serve in transport, storage, or membranes; or they may be toxins or enzymes. Their structures, like their functions, vary greatly. They are all, however, amino acid polymers arranged in a linear sequence.
A protein's shape is critical to its function. For example, an enzyme...
71.5K

You might also read

Related Articles

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

Sort by
Same author

Absorption Mode Broadband 2D MS for Proteomics and Metabolomics.

Journal of the American Society for Mass Spectrometry·2026
Same author

Exploring RNA G-Quadruplex Stability in the Gas Phase: Insights from Native Mass Spectrometry.

ChemPlusChem·2025
Same author

The intrinsic preference of guanosine bases for cleavage-facilitating interactions with phosphodiester moieties in RNA anions revealed by base modifications and mass spectrometry.

Nucleic acids research·2025
Same author

Engineering covalent small molecule-RNA complexes in living cells.

Nature chemical biology·2025
Same author

FAST MS: Software for the Automated Analysis of Top-Down Mass Spectra of Polymeric Molecules Including RNA, DNA, and Proteins.

Journal of the American Society for Mass Spectrometry·2024
Same author

Investigating the Intramolecular Competition of Different RNA Binding Motifs for Neomycin B by Native Top-Down Mass Spectrometry.

ChemPlusChem·2024

Related Experiment Video

Updated: May 4, 2026

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture
09:37

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

Published on: May 2, 2019

9.7K

Does Electron Capture Dissociation Cleave Protein Disulfide Bonds?

Barbara Ganisl1, Kathrin Breuker1

  • 1Institute for Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80-82, 6020 Innsbruck (Austria).

Chemistryopen
|December 24, 2013
PubMed
Summary

Mass spectrometry (MS) techniques like electron capture dissociation (ECD) and electron transfer dissociation (ETD) fragment peptides for analysis. New experimental data reveal that protein disulfide bond cleavage mechanisms in ECD/ETD are more complex than previously understood.

Keywords:
disulfide bondselectron capture dissociationmass spectrometryproteins

More Related Videos

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
07:16

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Published on: June 21, 2021

1.6K
High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli
12:16

High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli

Published on: July 30, 2014

26.3K

Related Experiment Videos

Last Updated: May 4, 2026

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture
09:37

Combining Non-reducing SDS-PAGE Analysis and Chemical Crosslinking to Detect Multimeric Complexes Stabilized by Disulfide Linkages in Mammalian Cells in Culture

Published on: May 2, 2019

9.7K
Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation
07:16

Resin-Assisted Capture Coupled with Isobaric Tandem Mass Tag Labeling for Multiplexed Quantification of Protein Thiol Oxidation

Published on: June 21, 2021

1.6K
High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli
12:16

High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli

Published on: July 30, 2014

26.3K

Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Proteomics

Background:

  • Mass spectrometry (MS) is crucial for peptide and protein characterization, relying on gas-phase dissociation to generate fragment ions.
  • Common methods like collision-induced dissociation (CID) cleave amide backbone bonds.
  • Electron-based methods, electron capture dissociation (ECD) and electron transfer dissociation (ETD), offer extensive sequence coverage via N-Cα bond cleavage and preserve labile post-translational modifications.

Purpose of the Study:

  • To investigate the mechanism of protein disulfide bond cleavage during ECD and ETD.
  • To experimentally validate or refute the postulate of preferential disulfide bond cleavage over backbone cleavage in electron-based dissociation methods.
  • To elucidate the intricate details of disulfide bond fragmentation in ECD/ETD.

Main Methods:

  • Utilized advanced mass spectrometry techniques, specifically electron capture dissociation (ECD) and electron transfer dissociation (ETD).
  • Performed experimental analysis of peptide and protein fragmentation patterns.
  • Focused on the cleavage of disulfide bonds in comparison to backbone bond cleavage.

Main Results:

  • Experimental data indicate that the mechanism of protein disulfide bond cleavage in ECD/ETD is significantly more complex than previously hypothesized.
  • The proposed preferential cleavage of disulfide bonds over backbone bonds in ECD/ETD is debated and requires further intricate mechanistic understanding.
  • The study provides new experimental evidence challenging existing models of disulfide bond fragmentation.

Conclusions:

  • The fragmentation mechanisms governing disulfide bond cleavage in electron-based mass spectrometry methods are intricate and not fully understood.
  • Existing postulates regarding preferential disulfide bond cleavage in ECD/ETD require re-evaluation based on new experimental findings.
  • Further research is necessary to comprehensively elucidate the complex interplay between electron capture/transfer, backbone cleavage, and disulfide bond rupture.