Crystal and Solution Structure Analysis of FhuD2 from <i>Staphylococcus aureus</i> in Multiple Unliganded Conformations
and Bound to Ferrioxamine‑B
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Abstract
Iron acquisition is a central process
for virtually all organisms.
In <i>Staphylococcus aureus</i>, FhuD2 is a lipoprotein
that is a high-affinity receptor for iron-bound hydroxamate siderophores.
In this study, FhuD2 was crystallized bound to ferrioxamine-B (FXB),
and also in its ligand-free state; the latter structures are the first
for hydroxamate-binding receptors within this protein family. The
structure of the FhuD2–FXB conformation shows that residues
W197 and R199 from the C-terminal domain donate hydrogen bonds to
the hydroxamate oxygens, and a ring of aromatic residues cradles the
aliphatic arms connecting the hydroxamate moieties of the siderophore.
The available ligand-bound structures of FhuD from <i>Escherichia
coli</i> and YfiY from <i>Bacillus cereus</i> show
that, despite a high degree of structural conservation, three protein
families have evolved with critical siderophore binding residues on
either the C-terminal domain (<i>S. aureus</i>), the N-terminal
domain (<i>E. coli</i>), or both (<i>B. cereus</i>). Unliganded FhuD2 was crystallized in five conformations related
by rigid body movements of the N- and C-terminal domains. Small-angle
X-ray scattering (SAXS) indicates that the solution conformation of
unliganded FhuD2 is more compact than the conformations observed in
crystals. The ligand-induced conformational changes for FhuD2 in solution
are relatively modest and depend on the identity of the siderophore.
The crystallographic and SAXS results are used to discuss roles for
the liganded and unliganded forms of FhuD2 in the siderophore transport
mechanism