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Fig. 3. Role of energy-coupled lipid flippases in triggering membrane budding. (A) ATP-driven lipid translocation may be required to generate a lipid imbalance across the bilayer by increasing the proportion of total lipids in one monolayer and thereby driving budding of vesicles. noL, number of lipid molecules in the outer leaflet, niL, number of lipid molecules in the inner leaflet (e.g. at the level of the head groups, the cytoplasmic leaflet of a 60 nm diameter vesicle contains 1.5 times the number of lipid molecules of the lumenal leaflet). (B) ATP-driven lipid translocation may help to create a high local concentration of aminophospholipids in the cytosolic leaflet favorable for recruitment of peripheral proteins, such as ARF, clathrin, amphiphysin and endophilins.