Formation de proteo-GUVs

Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies

Matthias Garten¹, Sophie Aimon², Patricia Bassereau¹, Gilman E. S. Toombes³

¹Institut Curie, Centre de Recherche, CNRS, UMR 168, PhysicoChimie Curie, Université Pierre et Marie Curie, ²Kavli Institute for Brain and Mind, University of California, San Diego, ³Molecular Physiology and Biophysics Section, National Institute for Neurological Disorders and Stroke, National Institute of Health

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The reconstitution of the transmembrane protein, KvAP, into giant unilamellar vesicles (GUVs) is demonstrated for two dehydration-rehydration methods — electroformation, and gel-assisted swelling. In both methods, small unilamellar vesicles containing the protein are fused together to form GUVs that can then be studied by fluorescence microscopy and patch-clamp electrophysiology.