4.5 Review

Native electrophoretic techniques to identify protein-protein interactions

Journal

PROTEOMICS
Volume 9, Issue 23, Pages 5214-5223

Publisher

WILEY
DOI: 10.1002/pmic.200900151

Keywords

Blue-native electrophoresis; Clear-native electrophoresis; Protein complex; Protein-protein interaction; Supercomplex; Technology

Funding

  1. Deutsche Forschungsgemeinschaft
  2. Sonderforschungsbereich [815]
  3. Cluster of Excellence Macromolecular Complexes at the Goethe University Frankfurt [EXC 115]

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Permanent protein-protein interactions are commonly identified by co-purification of two or more protein components using techniques like co-immunoprecipitation, tandem affinity purification and native electrophoresis. Here we focus on blue-native electrophoresis, clear-native electrophoresis, high-resolution clear-native electrophoresis and associated techniques to identify stable membrane protein complexes and detergent-labile physiological super-complexes. Hints for dynamic protein-protein interactions can be obtained using two-hybrid techniques but not from native electrophoresis and other protein isolation techniques except after covalent cross-linking of interacting proteins in vivo prior to protein separation.

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