Solubilization of membrane protein complexes for blue native PAGE.
Blue native PAGE is an electrophoretic technique for high-resolution separation of membrane proteins. The method has been proven especially useful for investigation of native protein complexes enabling a characterization of potential protein-protein interactions in the context of functional proteomics. Blue native PAGE is easy to realise, results are reproducible and a high number of protocols are available. However, care should be taken during solubilization of protein complexes to achieve significant results in BN-PAGE analysis. Solubilization of membranes and proteins is not only influenced by detergent-lipid and detergent-protein interactions but also by lipid-lipid, lipid-protein and protein-protein interactions. Interactions have been investigated experimentally and theoretically. But, in practice, the experimental results do not always mirror the theoretical basis and therefore optimal solubilization conditions for each membrane and membrane protein complex should be investigated individually to tap the full potential of BN-PAGE analysis.
Department Biology I, Ludwig-Maximilians-Universität München, Menzingerstrasse 67, Munich, Germany.
Veronika
V
Reisinger
Lutz Andreas
LA
Eichacker
D000818:Animals; D003902:Detergents / Q000737:chemistry / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005656:Fungal Proteins / Q000737:chemistry; D007477:Ions; D007525:Isoelectric Focusing / Q000379:methods; D008055:Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism; D025941:Protein Interaction Mapping; D020543:Proteome; D040901:Proteomics / Q000379:methods*
D003902:Detergents; D005656:Fungal Proteins; D007477:Ions; D008055:Lipids; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D020543:Proteome