Transfer of plasma membrane proteins between cells using reconstituted membrane vesicles as shuttle vehicles.
Purified membrane glycoproteins from liver or hepatoma tissue culture cells were incorporated in a right-side-out orientation into reconstituted phospholipid vesicles by a detergent dialysis method. The phospholipids were purified from membrane preparations of rat liver. The protein:phospholipid ratio of the reconstituted vesicles was optimized for efficient transfer of vesicle contents to the recipient cells, usually mouse L cells. Fluoresceinated albumin incorporated into the lumen of reconstituted vesicles was used as a marker for transfer after polyethylene glycol-mediated fusion. The redistribution and fate of both the lipids and the transferred membrane proteins were analysed by microscopic and biochemical methods. A hepatocyte-specific binding protein for galactose- or galactosamine-terminated serum glycoproteins and a set of hepatoma cell plasma membrane glycoproteins were successfully transferred to the plasma membrane of mouse fibroblasts by these methods. The biological function of the hepatic binding protein, namely delivery of the galactose-terminated glycoprotein ligand to the lysosome for degradation, was imparted to the mouse fibroblast after transfer. Further, both the polypeptide and the carbohydrate moieties of a set of membrane proteins were degraded at about the same relative rates as they had in the original donor cells, after transfer to the plasma membrane of recipient mouse fibroblasts. These studies show that the technique of inserting membrane constituents into the plasma membrane of another cell can help to elucidate the route and mechanism of membrane protein function and turnover.
D000818:Animals; D002459:Cell Fusion; D002462:Cell Membrane / Q000378:metabolism*; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D007739:L Cells / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D051381:Rats
D006023:Glycoproteins; D008565:Membrane Proteins