title: Role of Gly117 in the cation/melibiose symport of MelB of Salmonella typhimurium.
abstract: The melibiose permease of Salmonella typhimurium (MelB(St)) catalyzes symport of melibiose with Na(+), Li(+), or H(+), and bioinformatics analysis indicates that a conserved Gly117 (helix IV) is part of the Na(+)-binding site. We mutated Gly117 to Ala, Pro, Trp, or Arg; the effects on melibiose transport and binding of cosubstrates depended on the physical-chemical properties of the side chain. Compared with WT MelB(St), the Gly117 → Ala mutant exhibited little difference in either cosubstrate binding or stimulation of melibiose transport by Na(+) or Li(+), but all other mutations reduced melibiose active transport and efflux, and decreased the apparent affinity for Na(+). The bulky Trp at position 117 caused the greatest inhibition of melibiose binding, and Gly117 → Arg yielded less than a 4-fold decrease in the apparent affinity for melibiose at saturating Na(+) or Li(+) concentration. Remarkably, the mutant Gly117 → Arg catalyzed melibiose exchange in the presence of Na(+) or Li(+), but did not catalyze melibiose translocation involving net flux of the coupling cation, indicating that sugar is released prior to release of the coupling cation. Taken together, the findings are consistent with the notion that Gly117 plays an important role in cation binding and translocation.
authors.affiliation: Department of Cell Physiology & Molecular Biophysics, Center for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, USA. Lan.Guan@ttuhsc.edu
authors.forename: Lan
authors.initials: L
authors.lastname: Guan
authors.forename: S Vivek
authors.initials: SV
authors.lastname: Jakkula
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Hodkoff
authors.forename: Yue
authors.initials: Y
authors.lastname: Su
mesh_terms: D001426:Bacterial Proteins / Q000502:physiology*; D005998:Glycine / Q000502:physiology*; D008553:Melibiose / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D012486:Salmonella typhimurium / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D008553:Melibiose; D005998:Glycine