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New zwitterionic detergents improve the analysis of membrane proteins by two‐dimensional electrophoresis

Electrophoresis · 1998 · Vol. 19(11) · pp. 1901–1909

Mireille Chevallet(CEA Grenoble)Véronique Santoni(Centre National de la Recherche Scientifique)Alexandra Poinas(Institut de Biologie Structurale)David Rouquié(Institut des Sciences des Plantes de Montpellier)Alexandra Fuchs(Institut de Biologie Structurale)Sylvie Kieffer(Institut de Biologie et de Chimie des Protéines)Sylvie Kieffer(Institut de Biologie et de Chimie des Protéines)Joël Lunardi(CEA Grenoble)Jérôme Garin(Laboratoire de Chimie et Biologie des Métaux)Thierry Rabilloud✉(Commissariat à l'Énergie Atomique et aux Énergies Alternatives)

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Abstract

Severe quantitative loss of protein is often observed in high-resolution two-dimensional electrophoresis of membrane proteins, while the resolution is usually not affected. To improve the solubility of proteins in this technique, we tested denaturing cocktails containing various detergents and chaotropes. Best results were obtained with a denaturing solution containing urea, thiourea, and zwitterionic detergents, synthesized for this purpose. Among the dozen detergents synthesized and tested, amidosulfobetaines with an alkyl tail containing 14-16 carbons proved most efficient, solubilizing previously undetected membrane proteins.

Mass Spectrometry Techniques and Applications Analytical Chemistry and Chromatography Advanced Proteomics Techniques and ApplicationsChaotropic agentThioureaSolubilityChromatographyMembrane proteinUreaChemistryElectrophoresisResolution (logic)Membrane