RAPID PREPARATION OF GIANT UNILAMELLAR VESICLES

We report here a rapid evaporation method that produces in high yield giant unilamellar vesicles up to 50 mu m in diameter. The vesicles are obtained after only 2 min and can be prepared from different phosphol ipids, including L-alpha-phosphatidylcholine (lecithin), dipalmitoleoy l L-alpha-phosphatidylcholine, and beta-arachidonoyl gamma-palmitoyl L -alpha-phosphatidylcholine. Vesicles can be produced in distilled wate r and in Hepes, phosphate, and berate buffers in the pH range of 7.0 t o 11.5 with ionic strengths up to 50 mM. The short preparation time al lows encapsulation of labile molecular targets or enzymes with high ca talytic activities. Cell-sized proteoliposomes have been prepared in w hich gamma-glutamyltransferase (EC 2.3.2.2) was functionally incorpora ted into the membrane wall.