1887

Abstract

The bacterium , a swine pathogen, utilizes ferrichrome as an iron source. This study details the molecular cloning and sequencing of the genes involved in the uptake of this hydroxamate siderophore. Four ferric hydroxamate uptake () genes, , , and , were identified in a single operon, and these were found to encode proteins homologous to proteins of the systems of several bacteria, including . The gene encodes the 77 kDa outer-membrane protein (OMP) FhuA, the receptor for ferrichrome. FhuD is the 356 kDa periplasmic protein responsible for the translocation of ferric hydroxamate from the outer to the inner membrane. FhuC (285 kDa) and FhuB (694 kDa) are cytoplasmic-membrane-associated proteins that are components of an ABC transporter which internalizes the ferric hydroxamate. Reference strains of that represented serotypes 1 to 12 of this organism all tested positive for the four genes. When FhuA was affinity-tagged with hexahistidine at its amino terminus and expressed in an host, the recombinant protein reacted with an mAb against FhuA, as well as with a polyclonal pig serum raised against an infection. Hence, the authors conclude that is expressed by . Three-dimensional modelling of the OMP FhuA was achieved by threading it to the X-ray crystallographic structure of the homologous protein in . FhuA from was found to have the same overall fold as its homologue, i.e. it possesses an N-terminal cork domain followed by a C-terminal β-barrel domain and displays 11 extracellular loops and 10 periplasmic turns.

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2002-09-01
2019-10-18
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