1887

Abstract

The siderophore salmochelin is produced under iron-poor conditions by and many uropathogenic strains. The production of salmochelin, a C-glucosylated enterobactin, is dependent on the synthesis of enterobactin and the gene cluster. An IroD protein with an N-terminal His-tag cleaved cyclic salmochelin S4 to the linear trimer salmochelin S2, the dimer salmochelin S1, and the monomers dihydroxybenzoylserine and C-glucosylated dihydroxybenzoylserine (salmochelin SX, pacifarinic acid). The periplasmic IroE protein was purified as a MalE–IroE fusion protein. This enzyme degraded salmochelin S4 and ferric-salmochelin S4 to salmochelin S2 and ferric-salmochelin S2, respectively. In , uptake of ferric-salmochelin S4 was dependent on the cleavage by IroE, and independent of the FepBDGC ABC transporter in the cytoplasmic membrane. IroC, which has similarities to ABC-multidrug-resistance proteins, was necessary for the uptake of salmochelin S2 from the periplasm into the cytoplasm. IroE did not function as a classical binding protein since salmochelin S2 was taken up in the absence of a functional IroE protein. IroC mediated the uptake of iron via enterobactin in a mutant. IroE was also necessary in this case for the uptake of ferric-enterobactin, which indicated that only the linear degradation products of enterobactin were taken up via IroC. PfeE, the IroE homologue, was cloned, and its enzymic activity was shown to be very similar to that of IroE. It is suggested that homologues in other bacteria are also periplasmic IroE-type esterases of siderophores.

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2005-07-01
2019-10-23
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vol. , part 7, pp. 2363 - 2372

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Fig. S1. SDS-PAGE gels of the purified NHis-IroD fusion protein, the MalE-IroE fusion protein and the MalE-PfeE fusion protein. Fig. S2. Cleavage of salmochelin S2 by NHis-IroD. Fig. S3. Substrates that were not cleaved by MalE-IroE: Ga -salmochelin S4, Ga -enterobactin. Substrates that were not cleaved by NHis-IroD: Fe -enterobactin, Ga -salmochelin S4, Fe -salmochelin S4, salmochelin S0. Fig. S4. Cleavage of Fe -salmochelin S4 by NHis-IroD in the presence of 2 mM ferrozine. Fig. S5. Cleavage of enterobactin by NHis-IroD to the linear oligomers 2,3-dihydroxybenzoylserine. Fig. S6. Cleavage of salmochelin S4 by MalE-PfeE from . Fig. S7. Cleavage of Fe -salmochelin S4 by MalE-PfeE from .



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