A sensitive method to introduce membrane-bound proteins into recipient cells based on affinity enrichment of lipid vesicles to the recipient cell prior to fusion.
A sensitive analytical procedure for studying membrane-bound structures has been developed. Membrane glycoproteins inserted into liposomes were transferred to recipient cells by use of a lectin, concanavalin A, bound to the cells as a bridge to generate proximity between the recipient cell and the glycoprotein-containing liposome, prior to exposure to the fusing agent, poly(ethylene glycol). Partially purified histocompatibility antigen from rats was introduced into the membrane of human lymphocytes. After treating the cells with poly(ethylene glycol) under fusion conditions, some of the antigen present in the preparation could not be eluted with alpha-methyl mannoside and EDTA, indicating that incorporation in the cell membrane had taken place. This antigen remained exposed on the lymphocyte surface for approximately 1 h as demonstrated by sensitivity of the lymphocytes to the lytic effect of an antiserum to the histocompatibility antigen in the presence of complement. Some of the lectin molecules seemed to be internalized in the cells but no induction of cell mitosis was observed. The described method gives an opportunity to work with small amounts of membrane proteins inserted into liposomes, introducing them into recipient cells for analysis of their biological activities.
D000818:Animals; D002462:Cell Membrane / Q000276:immunology; D003208:Concanavalin A; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006017:Glycolipids; D006649:Histocompatibility Antigens; D006801:Humans; D008081:Liposomes; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D010714:Phosphatidylethanolamines; D051381:Rats; D011920:Rats, Inbred WF; D013154:Spleen / Q000276:immunology; D013861:Thiocyanates
D005452:Fluoresceins; D005456:Fluorescent Dyes; D006017:Glycolipids; D006649:Histocompatibility Antigens; D008081:Liposomes; D008565:Membrane Proteins; D010714:Phosphatidylethanolamines; D013861:Thiocyanates; C038353:histocompatibility antigens RT, rat; D003208:Concanavalin A; D016650:Fluorescein-5-isothiocyanate