Native electrophoretic techniques to identify protein-protein interactions

被引:71
|
作者
Wittig, Ilka [1 ]
Schaegger, Hermann [1 ]
机构
[1] Goethe Univ Frankfurt, Zentrum Biol Chem, Frankfurt, Germany
关键词
Blue-native electrophoresis; Clear-native electrophoresis; Protein complex; Protein-protein interaction; Supercomplex; Technology; POLYACRYLAMIDE-GEL ELECTROPHORESIS; OXIDATIVE-PHOSPHORYLATION COMPLEXES; TRANSPORT CHAIN COMPLEXES; CYTOCHROME-C-OXIDASE; DIMERIC ATP SYNTHASE; MEMBRANE-PROTEIN; FUNCTIONAL ASSAYS; HUMAN-DISEASES; YEAST; SUPERCOMPLEX;
D O I
10.1002/pmic.200900151
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:5214 / 5223
页数:10
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