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A simplified procedure for the isolation of bacterial polypeptide elongation factor EF-Tu

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Leberman,  Reuben
Max Planck Institute for Medical Research, Max Planck Society;

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Giovanelli,  Ruth
Max Planck Institute for Medical Research, Max Planck Society;

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Guariguata,  Rafael A.
Max Planck Institute for Medical Research, Max Planck Society;

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Schumann,  Renate
Dietmar Manstein Group, Max Planck Institute for Medical Research, Max Planck Society;

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Wittinghofer,  Alfred
Emeritus Group Biophysics, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

Leberman, R., Antonsson, B., Giovanelli, R., Guariguata, R. A., Schumann, R., & Wittinghofer, A. (1980). A simplified procedure for the isolation of bacterial polypeptide elongation factor EF-Tu. Analytical Biochemistry, 104(1), 29-36. doi:10.1016/0003-2697(80)90272-9.


Cite as: http://hdl.handle.net/21.11116/0000-0004-B383-D
Abstract
Polypeptide elongation factor EF-Tu can be isolated from bacterial cell extracts in two fractionation steps. The first is ion-exchange chromatography on DEAE-Sepharose, CL-6B, and the second is gel filtration on AcA 44. The method is illustrated with extracts from Escherichia coli, Bacillus stearothermophilus, and the thermophilic bacterium PS3. The extracts were obtained from lysozyme-treated cells and were processed without high-speed centrifugation or ammonium sulfate fractionation. The procedure is simple and rapid, gives higher yields than previous methods, and is easily scaled to any size preparation. The procedure also produces fractions enriched in the other polypeptide elongation factors EF-Ts and EF-G.