Towards structural studies of the old yellow enzyme homologue SYE4 from Shewanella oneidensis and its complexes at atomic resolution

1. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jan 1;66(Pt 1):85-90.
doi: 10.1107/S1744309109050386. Epub 2009 Dec 25.

Towards structural studies of the old yellow enzyme homologue SYE4 from
Shewanella oneidensis and its complexes at atomic resolution.

Elegheert J(1), van den Hemel D, Dix I, Stout J, Van Beeumen J, Brigé A,
Savvides SN.

Author information:
(1)Department of Biochemistry and Microbiology, Laboratory for Protein
Biochemistry and Biomolecular Engineering (L-ProBE), Ghent University, K. L.
Ledeganckstraat 35, 9000 Ghent, Belgium.

Shewanella oneidensis is an environmentally versatile Gram-negative
gamma-proteobacterium that is endowed with an unusually large proteome of redox
proteins. Of the four old yellow enzyme (OYE) homologues found in S. oneidensis,
SYE4 is the homologue most implicated in resistance to oxidative stress. SYE4
was recombinantly expressed in Escherichia coli, purified and crystallized using
the hanging-drop vapour-diffusion method. The crystals belonged to the
orthorhombic space group P2(1)2(1)2(1) and were moderately pseudo-merohedrally
twinned, emulating a P422 metric symmetry. The native crystals of SYE4 were of
exceptional diffraction quality and provided complete data to 1.10 A resolution
using synchrotron radiation, while crystals of the reduced enzyme and of the
enzyme in complex with a wide range of ligands typically led to high-quality
complete data sets to 1.30-1.60 A resolution, thus providing a rare opportunity
to dissect the structure-function relationships of a good-sized enzyme (40 kDa)
at true atomic resolution. Here, the attainment of a number of experimental
milestones in the crystallographic studies of SYE4 and its complexes are
reported, including isolation of the elusive hydride-Meisenheimer complex.

DOI: 10.1107/S1744309109050386
PMCID: PMC2805545
PMID: 20057079 [Indexed for MEDLINE]

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