title: Cation-pi interactions as determinants for binding of the compatible solutes glycine betaine and proline betaine by the periplasmic ligand-binding protein ProX from Escherichia coli.
abstract: Compatible solutes such as glycine betaine and proline betaine are accumulated to exceedingly high intracellular levels by many organisms in response to high osmolarity to offset the loss of cell water. They are excluded from the immediate hydration shell of proteins and thereby stabilize their native structure. Despite their exclusion from protein surfaces, the periplasmic ligand-binding protein ProX from the Escherichia coli ATP-binding cassette transport system ProU binds the compatible solutes glycine betaine and proline betaine with high affinity and specificity. To understand the mechanism of compatible solute binding, we determined the high resolution structure of ProX in complex with its ligands glycine betaine and proline betaine. This crystallographic study revealed that cation-pi interactions between the positive charge of the quaternary amine of the ligands and three tryptophan residues forming a rectangular aromatic box are the key determinants of the high affinity binding of compatible solutes by ProX. The structural analysis was combined with site-directed mutagenesis of the ligand binding pocket to estimate the contributions of the tryptophan residues involved in binding.
authors: Fachbereich Biologie, Universität Konstanz, Universitätsstrasse 10, D-78457 Konstanz, Germany.
authors: André
authors: A
authors: Schiefner
authors: Jason
authors: J
authors: Breed
authors: Linda
authors: L
authors: Bösser
authors: Susanne
authors: S
authors: Kneip
authors: Jutta
authors: J
authors: Gade
authors: Gudrun
authors: G
authors: Holtmann
authors: Kay
authors: K
authors: Diederichs
authors: Wolfram
authors: W
authors: Welte
authors: Erhard
authors: E
authors: Bremer
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001622:Betaine / Q000737:chemistry*; D001665:Binding Sites; D001671:Biochemistry / Q000379:methods*; D002244:Carbon; D002412:Cations; D018360:Crystallography, X-Ray; D004220:Disulfides; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry*; D008024:Ligands; D026901:Membrane Transport Proteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D033902:Periplasmic Binding Proteins / Q000737:chemistry*; D011392:Proline / Q000031:analogs & derivatives* / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014364:Tryptophan / Q000737:chemistry
chemical_list: D002412:Cations; D004220:Disulfides; D029968:Escherichia coli Proteins; D008024:Ligands; D026901:Membrane Transport Proteins; C053437:PROX protein, E coli; D033902:Periplasmic Binding Proteins; D001622:Betaine; D002244:Carbon; D014364:Tryptophan; D011392:Proline; C003342:stachydrine