Binding of iron-free siderophore, a common feature of siderophore outer membrane transporters of Escherichia coli and Pseudomonas aeruginosa.
TonB-dependent iron transporters present in the outer membranes of Gram-negative bacteria transport ferric-siderophore complexes into the periplasm. This requires proton motive force and an integral inner membrane complex, TonB-ExbB-ExbD. Recognition of iron-free siderophores by TonB-dependent outer membrane transporters (OMT) has only been described for a subfamily called OMT(N). These OMT(N)s have an additional domain at the N terminus, which interacts with an inner membrane regulatory protein to activate a cytoplasmic sigma factor. This induces transcription of iron transport genes. Here we showed that the ability to bind aposiderophores is not specific to the OMT(N) subfamily but may be a more general feature of OMTs. FhuA, the ferrichrome OMT in Escherichia coli, and FptA, the pyochelin (Pch) OMT in Pseudomonas aeruginosa, were both able to bind in vitro and in vivo the apo-forms and the ferric forms of their corresponding siderophore at a common binding site. FptA produced in P. aeruginosa cells grown in an iron-deficient medium copurifies with a ligand that, as characterized by fluorescence, is iron-free Pch. As described previously for the FpvA transporter (pyoverdine OMT in P. aeruginosa), it appears that in conditions of iron limitation all the FptA receptors at the cell surface are loaded with apoPch. This FptA-Pch complex is less stable in vitro than the previously described copurified FpvA-Pvd complex and can be loaded with iron in vitro in the presence of Pch-Fe, citrate-Fe, or ferrichrome-Fe. These findings improved our understanding of the iron uptake mechanism via siderophores in Gram-negative bacteria.
Hoegy
Françoise
F
Département des Récepteurs et Protéines Membranaires, UPR 9050 CNRS, ESBS, Bld. Sébastien Brant, F-67413 Illkirch, Strasbourg, France.
Celia
Hervé
H
Mislin
Gaëtan L
GL
Vincent
Michel
M
Gallay
Jacques
J
Schalk
Isabelle J
IJ
D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000502:physiology; D001692:Biological Transport; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D005290:Ferric Compounds / Q000378:metabolism; D005291:Ferrichrome / Q000378:metabolism; D005453:Fluorescence; D005454:Fluorescence Polarization; D007501:Iron / Q000378:metabolism*; D008565:Membrane Proteins / Q000502:physiology; D010636:Phenols / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D011991:Receptors, Virus / Q000378:metabolism*; D017262:Siderophores / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013844:Thiazoles / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C075523:FPTA protein, Pseudomonas; D005290:Ferric Compounds; C050900:FhuA protein, E coli; D008565:Membrane Proteins; D010636:Phenols; D011956:Receptors, Cell Surface; D011991:Receptors, Virus; D017262:Siderophores; D013844:Thiazoles; C035902:siderophore receptors; C057616:tonB protein, Bacteria; D005291:Ferrichrome; C025316:pyochelin; D007501:Iron