A rapid reverse phase evaporation method for the reconstitution of uncharged thylakoid membrane lipids that resist hydration

Comparison of several lipid reconstitution methods showed that they were not equally efficient at transferring the predominant thylakoid lipid, monogalactosyldiglyceride (MG), to the aqueous phase. We report a reverse phase evaporation method that employs Freon 11 as a lipid solvent and is capable of successfully hydrating MG in spinach (Spinacia oleracea L.) at room temperature within minutes. Using this method it is possible to force an equal weight mixture of MG and digalactosyldiglyceride into small bilayer vesicles without the formation of inverted micellar `lipidic particles' in the membranes.