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

Membrane binding of a cyanobacterial ESCRT-III protein crucially involves the helix α1-3 hairpin conserved in all superfamily members.

Protein science : a publication of the Protein Society·2025
Same author

Direct interaction between TDP-43 and Tau promotes their co-condensation, while suppressing Tau fibril formation and seeding.

The EMBO journal·2025
Same author

Age-related changes in the proteome and mitochondrial metabolism of rabbit adipose-derived stromal/stem cells.

Scientific reports·2025
Same author

Effects of Surface Charge of Amphiphilic Peptides on Peptide-Lipid Interactions in the Gas Phase and in Solution.

Analytical chemistry·2025
Same author

MS <i>SIEVE</i>-Pushing the Limits for Biomolecular Mass Spectrometry.

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

Nutrient Intakes in Prostate Cancer Survivors in the United States: A Nationally Representative Study.

Nutrition and cancer·2024

Related Experiment Video

Updated: Apr 25, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

20.6K

A comparative cross-linking strategy to probe conformational changes in protein complexes.

Carla Schmidt1, Carol V Robinson1

  • 1Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford, UK.

Nature Protocols
|August 22, 2014
PubMed
Summary

This study introduces a comparative cross-linking method using mass spectrometry (MS) to detect protein conformational changes. The technique quantifies alterations in subunit interactions upon stimuli like ligand binding or post-translational modifications.

More Related Videos

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
12:43

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study

Published on: July 27, 2016

12.6K
Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
11:33

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking

Published on: December 17, 2013

5.8K

Related Experiment Videos

Last Updated: Apr 25, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
10:01

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

Published on: November 28, 2017

20.6K
Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study
12:43

Genetic and Biochemical Approaches for In Vivo and In Vitro Assessment of Protein Oligomerization: The Ryanodine Receptor Case Study

Published on: July 27, 2016

12.6K
Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
11:33

Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking

Published on: December 17, 2013

5.8K

Area of Science:

  • Biochemistry
  • Proteomics
  • Structural Biology

Background:

  • Chemical cross-linking with mass spectrometry (MS) is vital for studying static protein assemblies.
  • Probing dynamic conformational changes in protein complexes requires advanced methodologies.

Purpose of the Study:

  • To develop and validate a comparative cross-linking strategy for analyzing stimulus-induced conformational changes in protein complexes.
  • To quantify alterations in protein subunit interactions in response to various biological stimuli.

Main Methods:

  • Utilized lysine-specific deuterated and nondeuterated bis(sulfosuccinimidyl)suberate (BS3) cross-linking reagents.
  • Employed a comparative approach involving cross-linking of protein complexes with and without specific stimuli (ligand binding, post-translational modifications).
  • Analyzed cross-linked peptides using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database searching, comparing labeled and unlabeled ratios.

Main Results:

  • Successfully quantified changes in subunit interactions of ATP synthase induced by dephosphorylation.
  • Demonstrated the method's ability to detect conformational shifts triggered by ligand binding and other post-translational modifications.
  • Established a protocol applicable to various protein complexes and stimuli.

Conclusions:

  • The developed comparative cross-linking strategy provides a robust method for investigating dynamic protein complex behavior.
  • This technique offers a direct readout of stimulus-induced conformational changes, advancing the understanding of protein function.
  • The protocol is efficient, with application taking approximately 9 days, including purification.