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Native electrophoretic techniques to identify protein–protein interactions
Author(s) -
Wittig Ilka,
Schägger Hermann
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200900151
Subject(s) - electrophoresis , immunoprecipitation , protein–protein interaction , protein tag , protein purification , tandem affinity purification , gel electrophoresis of proteins , membrane protein , affinity electrophoresis , gel electrophoresis , biochemistry , proteomics , biology , chemistry , polyacrylamide gel electrophoresis , chromatography , affinity chromatography , enzyme , recombinant dna , membrane , fusion protein , gene
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 supercomplexes. 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.