title: Improved technique for reconstituting incredibly high and soluble amounts of tetrameric K⁺ channel in natural membranes.
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.
authors.affiliation: Institute for Biophysics, Johannes Kepler University Linz, 4040 Linz, Austria. mraja@pmcol.ualberta.ca
authors.forename: Mobeen
authors.initials: M
authors.lastname: Raja
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Vales
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D057005:Bioengineering / Q000379:methods*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002793:Cholic Acids / Q000737:chemistry; D003001:Cloning, Molecular; D003902:Detergents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005960:Glucosides / Q000737:chemistry; D010957:Plasmids; D015221:Potassium Channels / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D055503:Protein Multimerization; D011510:Proteolipids / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D002793:Cholic Acids; D003902:Detergents; D005960:Glucosides; D015221:Potassium Channels; D011510:Proteolipids; D011994:Recombinant Proteins; C020485:proteoliposomes; C018619:octyl-beta-D-glucoside; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate