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Two-dimensional electrophoresis of membrane proteins

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Abstract

One third of all genes of various organisms encode membrane proteins, emphasizing their crucial cellular role. However, due to their high hydrophobicity, membrane proteins demonstrate low solubility and a high tendency for aggregation. Indeed, conventional two-dimensional gel electrophoresis (2-DE), a powerful electrophoretic method for the separation of complex protein samples that applies isoelectric focusing (IEF) in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension, has a strong bias against membrane proteins. This review describes two-dimensional electrophoretic techniques that can be used to separate membrane proteins. Alternative methods for performing conventional 2-DE are highlighted; these involve replacing the IEF with electrophoresis using cationic detergents, namely 16-benzyldimethyl-n-hexadecylammonium chloride (16-BAC) and cetyl trimethyl ammonium bromide (CTAB), or the anionic detergent SDS. Finally, the separation of native membrane protein complexes through the application of blue and clear native gel electrophoresis (BN/CN-PAGE) is reviewed, as well as the free-flow electrophoresis (FFE) of membranes.

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Fig. 1 a–e
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Acknowledgments

We wish to especially thank J. Bucher for excellent technical assistance. This work was funded by the European Union EU-IP EVI-GENORET (LSHG-CT-2005-512036) and EU-IP Interaction Proteome (LSHG-CT-2003-505520), and by the German Federal Ministry of Education and Research (BMBF) NGFN2 SMP Proteomics (FKZ: 01GR0449, subproject 9) and QuantPro (FKZ: 0313865A, subproject A).

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  1. GSF–National Research Center for Environment and Health, Institute of Human Genetics, Ingolstaedter Landstrasse 1, 85764, Munich–Neuherberg, Germany

    Ralf J. Braun, Norbert Kinkl, Monika Beer & Marius Ueffing

  2. Technical University Munich, Institute of Human Genetics, 81675, Munich, Germany

    Norbert Kinkl & Marius Ueffing

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  1. Ralf J. Braun
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Correspondence to Marius Ueffing.

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Braun, R.J., Kinkl, N., Beer, M. et al. Two-dimensional electrophoresis of membrane proteins. Anal Bioanal Chem 389, 1033–1045 (2007). https://doi.org/10.1007/s00216-007-1514-6

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