Enhanced detergent extraction for analysis of membrane proteomes by two-dimensional gel eletrophoresis

被引：55

作者：

Churchward M.A. ^{[1
]}

Butt R.H. ^{[1
]}

Lang J.C. ^{[1
]}

Hsu K.K. ^{[1
]}

Coorssen J.R. ^{[1
,2
,3
]}

机构：

[1] Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1

[2] Dept. of Biochemistry and Molecular Biology, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1

[3] Hotchkiss Brain Institute, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1

来源：

Proteome Science | / 3卷 / 1期

关键词：

Cantharidin; Total Protein Assay; Zwitterionic Detergent; Monododecyl Ether; Total Detergent;

D O I：

10.1186/1477-5956-3-5

中图分类号：

学科分类号：

摘要：

Background: The analysis of hydrophobic membrane proteins by two-dimensional gel electrophoresis has long been hampered by the concept of inherent difficulty due to solubility issues. We have optimized extraction protocols by varying the detergent composition of the solubilization buffer with a variety of commercially available non-ionic and zwitterionic detergents and detergent-like phospholipids. Results: After initial analyses by one-dimensional SDS-PAGE, quantitative two-dimensional analyses of human erythrocyte membranes, mouse liver membranes, and mouse brain membranes, extracted with buffers that included the zwitterionic detergent MEGA 10 (decanoyl-N-methylglucamide) and the zwitterionic lipid LPC (1-lauroyl lysophosphatidylcholine), showed selective improvement over extraction with the common 2-DE detergent CHAPS (3[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate). Mixtures of the three detergents showed additive improvements in spot number, density, and resolution. Substantial improvements in the analysis of a brain membrane proteome were observed. Conclusions: This study demonstrates that an optimized detergent mix, coupled with rigorous sample handling and electrophoretic protocols, enables simple and effective analysis of membrane proteomes using two-dimensional electrophoresis. © 2005 Churchward et al., licensee BioMed Central Ltd.

引用

页数：11

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