Solubilization and reconstitution of membrane proteins of Escherichia coli using alkanoyl-N-methylglucamides.
Alkanoyl-N-methylglucamides, nonionic detergents, were utilized to solubilize membrane proteins of Escherichia coli and were used to reconstitute them into liposomes. First, critical micelle concentrations (CMC) of nonanoyl-N-methylglucamide and decanoyl-N-methylglucamide were determined to be 25 mM and 7 mM, respectively, by photometric assay. Then solubilization and reconstitution of the melibiose transport carrier were performed using these detergents at concentrations above the CMC. Melibiose counterflow activity was observed with the proteoliposomes reconstituted from the extracted proteins and phospholipids. The proton-translocating ATPase complex (F1-F0) was also solubilized with these detergents. These results indicate that nonanoyl- and decanoyl-N-methylglucamide are useful detergents for solubilization and reconstitution of membrane proteins.
D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003902:Detergents; D004926:Escherichia coli / Q000378:metabolism* / Q000648:ultrastructure; D005227:Fatty Acids; D005944:Glucosamine / Q000031:analogs & derivatives; D007700:Kinetics; D008081:Liposomes; D008553:Melibiose / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000302:isolation & purification; D012995:Solubility
D003902:Detergents; D005227:Fatty Acids; D008081:Liposomes; D008565:Membrane Proteins; C042222:nonanoyl-N-methylglucamide; C042221:decanoyl-N-methylglucamide; D008553:Melibiose; D006180:Proton-Translocating ATPases; D005944:Glucosamine