Journal
/
Biologie
/
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies

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

DOI:
10.3791/56747-v

•

10:01 min

•

November 28, 2017

•

, , , , ,
1Interdisciplinary research center HALOmem,Martin Luther University Halle-Wittenberg, 2Department of Chemistry,Kings College London

Chapitres

  • 00:05Titre
  • 00:51Mass Spectrometry-based Protein Identification
  • 04:35Mass Spectrometry of Intact Protein Complexes (Native Mass Spectrometry)
  • 06:17Chemical Cross-linking Coupled with Mass Spectrometry
  • 07:19Results: Chemical Cross-linking and Mass Spectrometry of Multi-subunit Protein Complexes
  • 09:14Conclusion

Summary

Traduction automatique

Traduction automatique

The architecture of protein complexes is essential for their function. Combining various mass spectrometric techniques proved powerful to study their assembly. We provide protocols for chemical cross-linking and native mass spectrometry and show how these complementary techniques help to elucidate the architecture of multi-subunit protein assemblies.

Tags

Chemical Cross-linkingMass SpectrometryIntact Protein ComplexesMulti-subunit Protein AssembliesStructural InformationCompositionConnectivityStoichiometryProtein Complex PurificationElectrophoresisCoomassie StainingProtein Band ExcisionDisulfide ReductionCysteine AlkylationTrypsin DigestionPeptide Extraction

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Isolation and Biophysical Study of Fruit Cuticles

Identification of Protein Complexes in Escherichia coli using Sequential Peptide Affinity Purification in Combination with Tandem Mass Spectrometry

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors

Identification of Post-translational Modifications of Plant Protein Complexes

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

Read Article