Abstract
The reconstitution of large amounts of integral proteins into lipid vesicles is largely prompted by the complexity of most biological membranes and protein stability. We optimized a particular system which maximized the incorporation efficiency of large soluble amounts of KcsA potassium channel in Escherichia coli membranes. The effects of two detergents, octylglucoside and 3-[(cholamidopropyl)-dimethyl-ammonio]-1-propanesulfonate (CHAPS), on KcsA reconstitution were compared. Reconstitution efficiency was found to be incredibly high for CHAPS-treated proteoliposomes followed by dialysis at room temperature. This approach may allow more accurate investigation of integral membrane proteins in their natural membrane environment via biophysical or biochemical techniques.
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Acknowledgements
This study was supported by the Federation of European Biochemical Societies (FEBS) and by funds from the Austrian Science Fund (FWF). We thank the late Sapar Saparov and the late Andreas Missner for their helpful suggestions and technical assistance in optimizing experimental conditions. The useful tips of Dr. Christopher Miller are gratefully acknowledged.
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Raja, M., Vales, E. Improved Technique for Reconstituting Incredibly High and Soluble Amounts of Tetrameric K+ Channel in Natural Membranes. J Membrane Biol 241, 141–144 (2011). https://doi.org/10.1007/s00232-011-9370-x
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DOI: https://doi.org/10.1007/s00232-011-9370-x