Improvement of two-dimensional gel electrophoresis proteome maps of the haloarchaeon Haloferax volcanii |
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Authors: | Karadzic Ivanka M Maupin-Furlow Julie A |
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Institution: | Department of Microbiology and Cell Science, University of Florida, Gainsville, FL 32611, USA. |
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Abstract: | Proteins of haloarchaea are remarkably unstable in low-ionic-strength solvents and tend to aggregate under standard two-dimensional (2-D) gel electrophoresis conditions, causing strong horizontal streaking. We have developed a new approach to generate 2-D maps of halophilic proteins which included washing cells with 1.5 M Tris-HCl buffer. In addition, proteins were precipitated with acetone, solubilized with urea and thiourea in the presence of the sulfobetaine detergent 3-(3-cholamidopropyl)dimethylamino]-1-propanesulfonate (CHAPS), reduced with tributylphosphine (TBP), and separated with microrange strips of immobilized pH gradients (pH 3.9-5.1). This combination enabled the construction of highly reproducible 2-D maps of Haloferax volcanii proteins. |
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Keywords: | Archaea Haloarchaea Halophile Proteome Two‐dimensional gel electrophoresis |
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