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Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment

Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment

DOI:
10.3791/57479-v

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09:02 min

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April 20, 2018

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,
1Cluster of excellence CellNetworks,Heidelberg University, 2Heidelberg University Biochemistry Center (BZH)

Chapters

  • 00:04Title
  • 00:46Split-BioID for Proteomics Studies
  • 02:54Streptavidin Pulldown
  • 05:03SDS-PAGE and Western Blotting
  • 06:21Results: Protein Expression and Biotinylation Through Split-BioID
  • 08:04Conclusion

Summary

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We provide a step-by-step protocol for split-BioID, a protein fragments-complementation assay based on the proximity-labeling technique BioID. Activated on the interaction of two given proteins, it allows the proteomics analysis of context-dependent protein complexes in their native cellular environment. The method is simple, cost-effective and only requires standard laboratory equipment.

Tags

Split-BioIDProteomic AnalysisProtein ComplexesNative Cellular EnvironmentProtein Fragment Complementation AssayProximity-dependent Protein BiotinylationCell BiologyMass Spectrometric AnalysisStreptavidin PulldownProtein Complexes RemodelingContext-specific Protein Complexes

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