Siderophores are iron chelators produced by bacteria to access iron， an essential nutriment. Pyoverdine (PVDI)， the major siderophore produced by Pseudomonas aeruginosa PAO1， consists of a fluorescent chromophore linked to an octapeptide. The ferric form of PVDI is transported from the extracellular environment into the periplasm by the outer membrane transporter， FpvA. Iron is then released from the siderophore in the periplasm by a mechanism that does not involve chemical modification of the chelator but an iron reduction step. Here， we followed the kinetics of iron release from PVDI， in vitro and in living cells， by monitoring its fluorescence (as apo PVDI is fluorescent， whereas PVDI-Fe(III)... More
Siderophores are iron chelators produced by bacteria to access iron， an essential nutriment. Pyoverdine (PVDI)， the major siderophore produced by Pseudomonas aeruginosa PAO1， consists of a fluorescent chromophore linked to an octapeptide. The ferric form of PVDI is transported from the extracellular environment into the periplasm by the outer membrane transporter， FpvA. Iron is then released from the siderophore in the periplasm by a mechanism that does not involve chemical modification of the chelator but an iron reduction step. Here， we followed the kinetics of iron release from PVDI， in vitro and in living cells， by monitoring its fluorescence (as apo PVDI is fluorescent， whereas PVDI-Fe(III) is not). Deletion of the inner membrane proteins fpvG (PA2403) and fpvH (PA2404) affected Fe uptake via PVDI and completely abolished PVDI-Fe dissociation， indicating that these two proteins are involved in iron acquisition via this siderophore. PVDI-Fe dissociation studies， using an in vitro assay， showed that iron release from this siderophore requires the presence of an iron reducer (DTT) and an iron chelator (ferrozine). In this assay， DTT could be replaced by the inner membrane protein， FpvG， and ferrozine by the periplasmic protein， FpvC， suggesting that FpvG acts as a reductase and FpvC as an Fe chelator in the process of PVDI-Fe dissociation in the periplasm of P. aeruginosa cells. This mechanism of iron release from PVDI is atypical among Gram-negative bacteria but seems to be conserved among Pseudomonads.
