title: The interaction between pyoverdin and its outer membrane receptor in Pseudomonas aeruginosa leads to different conformers: a time-resolved fluorescence study.
abstract: In iron limitation conditions, Pseudomonas aeruginosa secretes a major fluorescent siderophore named pyoverdin (PaA). PaA has an extremely high affinity for Fe(3+) but also chelates other ions such as Al(3+) and Ga(3+) with a lower affinity. The transfer of PaA-Fe(3+) across the outer membrane of the bacteria is mediated by the receptor FpvA, a TonB-dependent outer membrane transport protein. FpvA binds the iron-free and iron-loaded forms of pyoverdin with similar affinities, but only PaA-Fe(3+) is taken up by the cell, suggesting that FpvA adopts different conformations depending on its loading status. We used time-resolved fluorescence spectroscopy to characterize the different forms of FpvA-PaA in vitro. We showed that the FpvA-PaA complex adopts two different conformations depending on how it was prepared (formed in vitro or in vivo prior to purification). The dihydroquinoline moiety of both conformers is fully protonated, or coordinated by protein charged groups, but the polarity of its environment, its solvent accessibility, and its rotational dynamics are much slower when the FpvA-PaA complex is formed in vivo than in vitro. In the presence of Ga(3+) or Al(3+) ions, the solvent accessibility and mobility of the dihydroquinoline moiety in the two FpvA-PaA complexes are intermediate between those observed for the metal-free ones. In addition, the Förster resonance energy transfer kinetics from FpvA tryptophan residues to the PaA chromophore differs from one complex to the other, revealing differences in one or more of the donor-acceptor topologies.
authors.affiliation: ESBS, Ecole Supérieure de Biotechnologie de Strasbourg, UPR CNRS 9050, Bld Sébastien Brant, 67400 Illkirch, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Folschweiller
authors.forename: Jacques
authors.initials: J
authors.lastname: Gallay
authors.forename: Michel
authors.initials: M
authors.lastname: Vincent
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Abdallah
authors.forename: Franc
authors.initials: F
authors.lastname: Pattus
authors.forename: Isabelle J
authors.initials: IJ
authors.lastname: Schalk
mesh_terms: D000535:Aluminum / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D003902:Detergents / Q000737:chemistry; D005454:Fluorescence Polarization / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005708:Gallium / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D009842:Oligopeptides; D010860:Pigments, Biological / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000378:metabolism*; D017262:Siderophores / Q000737:chemistry; D012995:Solubility; D012996:Solutions; D013696:Temperature; D014867:Water / Q000737:chemistry
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; C082326:FpvA protein, Pseudomonas aeruginosa; D046911:Macromolecular Substances; D009842:Oligopeptides; D010860:Pigments, Biological; D017262:Siderophores; D012996:Solutions; D014867:Water; C042453:pyoverdin; D005708:Gallium; D000535:Aluminum