CARBOXYACYL DERIVATIVES OF CARDIOLIPIN AS 4-TAILED HYDROPHOBIC ANCHORS FOR THE COVALENT COUPLING OF HYDROPHILIC PROTEINS TO LIPOSOMES

Two carboxyacyl derivatives of cardiolipin, O-succinyl- and O-glutarylcardiolipin, were synthesized with the aim of using them as artificial membrane anchors for the immobilization of hydrophilic proteins to liposomes. Four adjacent fatty acid residues can be introduced into a protein with only one single amino group being blocked, by reacting the cardiolipin derivatives with the protein amino groups after carbodiimide activation. Alpha-chymotrypsin, used as a model protein, and modified with on average two molecules of O-succinylcardiolipin was incorporated into liposomes, which had been prepared by different methods, with very high yield. If incorporated in preformed liposomes, the carboxyacyl cardiolipin anchors were also efficient in binding proteins to liposomal surfaces. Up to 350-mu-g chymotrypsin/mu-mol lipid were coupled to small unilamellar vesicles, preserving reactivity of the enzyme towards specific macromolecular inhibitors. Human IgG could also be bound to anchor-containing liposomes with high protein to lipid coupling ratio as well as high coupling yield.