Reconstitution and function of membrane proteins into asymmetric giant liposomes by using a pulsed jet flow

This report describes preliminary results of a reconstitution and function of membrane proteins (flippases) into asymmetric lipid giant liposomes (GLs) that were prepared by deforming a planar asymmetric bilayer using a pulsed jet flow. Frist, we confirmed the expression of flippases on the baculovirus membrane by a western blot analysis. Next, we successfully reconstituted flippases into the asymmetric GL of phosphatidylserine by a baculovirus-liposome fusion method. Finally, we observed the translocation of phosphatidylserine from the extracellular leaflet to the cytoplasmic leaflet which was catalyzed by flippases. The flippase-reconstituted asymmetric liposomes that emulate the function of cell membranes will be useful for the study of elementary lipid-lipid or lipid-protein interactions.