title: Structure-based design of potent small-molecule binders to the S-component of the ECF transporter for thiamine.
abstract: Energy-coupling factor (ECF) transporters are membrane-protein complexes that mediate vitamin uptake in prokaryotes. They bind the substrate through the action of a specific integral membrane subunit (S-component) and power transport by hydrolysis of ATP in the three-subunit ECF module. Here, we have studied the binding of thiamine derivatives to ThiT, a thiamine-specific S-component. We designed and synthesized derivatives of thiamine that bind to ThiT with high affinity; this allowed us to evaluate the contribution of the functional groups to the binding affinity. We determined six crystal structures of ThiT in complex with our derivatives. The structure of the substrate-binding site in ThiT remains almost unchanged despite substantial differences in affinity. This work indicates that the structural organization of the binding site is robust and suggests that substrate release, which is required for transport, requires additional changes in conformation in ThiT that might be imposed by the ECF module.
authors: Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen (The Netherlands) http://www.rug.nl/research/membrane-enzymology/
authors: Lotteke J Y M
authors: LJ
authors: Swier
authors: Leticia
authors: L
authors: Monjas
authors: Albert
authors: A
authors: Guskov
authors: Alrik R
authors: AR
authors: de Voogd
authors: Guus B
authors: GB
authors: Erkens
authors: Dirk J
authors: DJ
authors: Slotboom
authors: Anna K H
authors: AK
authors: Hirsch
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001692:Biological Transport; D015195:Drug Design; D013294:Lactococcus lactis / Q000737:chemistry; D008958:Models, Molecular; D015394:Molecular Structure; D054852:Small Molecule Libraries / Q000737:chemistry / Q000378:metabolism*; D013831:Thiamine / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D054852:Small Molecule Libraries; D013831:Thiamine
keywords: X-ray diffraction; energy-coupling factor transporter; membrane proteins; molecular recognition; structure-based design