Thiolation of preformed liposomes with iminothiolane

Preformed phosphatidylethanolamine-containing liposomes were thiolated with 2-iminothiolane (Traut's reagent) and subsequently activated by mixed disulfide formation with 5,5'-dithiobis(2-nitrobenzoic acid). Up to 65% of amino groups of the outer liposomal lamella, corresponding to 230 SH-groups per vesicle, were modified. Covalent attachment of thiolated alpha-chymotrypsin to these thiol-liposomes via S-S bridges yielded a protein/lipid ratio of 3.6 X 10(-4) mol enzyme/mol lipid.