부산대학교 도서관

Simple Summary: Scramblases are proteins that translocate phospholipids from one leaflet of a membrane bilayer to the other. Our work identifies that a well-characterized protein from Escherichia coli, lactose permease, also displays lipid scrambling activity. The scrambling function is independent of LacY's proton-coupled lactose transport activity and involves two amino acid residues located near the membrane surface. Lactose permease (LacY) from Escherichia coli belongs to the major facilitator superfamily. It facilitates the co-transport of β-galactosides, including lactose, into cells by using a proton gradient towards the cell. We now show that LacY is capable of scrambling glycerophospholipids across a membrane. We found that purified LacY reconstituted into liposomes at various protein to lipid ratios catalyzed the rapid translocation of fluorescently labeled and radiolabeled glycerophospholipids across the proteoliposome membrane bilayer. The use of LacY mutant proteins unable to transport lactose revealed that glycerophospholipid scrambling was independent of H+/lactose transport activity. Unexpectedly, in a LacY double mutant locked into an occluded conformation glycerophospholipid, scrambling activity was largely inhibited. The corresponding single mutants revealed the importance of amino acids G46 and G262 for glycerophospholipid scrambling of LacY. [ABSTRACT FROM AUTHOR]