Structure-based mechanism for Na+/melibiose symport by MelB

The bacterial melibiose permease (MelB) belongs to the glycoside–pentoside–hexuronide:cation symporter family, a part of the major facilitator superfamily (MFS). Structural information regarding glycoside–pentoside–hexuronide:cation symporter family transporters and other Na+-coupled permeases within MFS has been lacking, although a wealth of biochemical and biophysical data are available. Here we present the three-dimensional crystal structures of Salmonella typhimurium MelBSt in two conformations, representing an outward partially occluded and an outward inactive state of MelBSt. MelB adopts a typical MFS fold and contains a previously unidentified cation-binding motif. Three conserved acidic residues form a pyramidal-shaped cation-binding site for Na+, Li+ or H+, which is in close proximity to the sugar-binding site. Both cosubstrate-binding sites are mainly contributed by the residues from the amino-terminal domain. These two structures and the functional data presented here provide mechanistic insights into Na+/melibiose symport. We also postulate a structural foundation for the conformational cycling necessary for transport catalysed by MFS permeases in general. The bacterial symporter MelB transports galactosides with Na+, Li+ or H+ ions. Ethayathulla et al. present crystal structures of MelB in two distinct conformations, providing the first structural insights into the transport mechanism of a sodium-coupled permease of the major facilitator superfamily.