Energetics of lipid transport by the ABC transporter MsbA is lipid dependent

The ABC multidrug exporter MsbA mediates the translocation of lipopolysaccharides and phospholipids across the plasma membrane in Gram-negative bacteria. Although MsbA is structurally well characterised, the energetic requirements of lipid transport remain unknown. Here, we report that, similar to the transport of small-molecule antibiotics and cytotoxic agents, the flopping of physiologically relevant long-acyl-chain 1,2-dioleoyl (C18)-phosphatidylethanolamine in proteoliposomes requires the simultaneous input of ATP binding and hydrolysis and the chemical proton gradient as sources of metabolic energy. In contrast, the flopping of the large hexa-acylated (C12-C14) Lipid-A anchor of lipopolysaccharides is only ATP dependent. This study demonstrates that the energetics of lipid transport by MsbA is lipid dependent. As our mutational analyses indicate lipid and drug transport via the central binding chamber in MsbA, the lipid availability in the membrane can affect the drug transport activity and vice versa. Guo et al. conduct a biochemical analysis of the energy dependence of substrate transport by MsbA, an essential ABC lipid transporter in Gram-negative bacteria. The authors examine the flopping of physiologically relevant lipids by MsbA and their differing dependencies on ATP and chemical proton gradient, demonstrating that the energetics of lipid transport by MsbA is lipid dependent.