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

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10:01 min

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November 28, 2017

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

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.

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

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