SOLVING PHASE PROBLEM USING A SE-MET DERIVATIVE OF THE FLAVOENZYME NAD(P)H:ACCEPTOR OXIDOREDUCTASE (FERB)

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Authors

KLUMPLER Tomáš MAREK Jaromír SEDLÁČEK Vojtěch KUČERA Igor

Year of publication 2010
Type Article in Proceedings
Conference Materials Structure, vol. 17, no. 1a (2010)
MU Faculty or unit

Faculty of Science

Citation
Field Biochemistry
Keywords phase problem; flavoenzyme; FerB; reductase; crystallography
Description The flavin adenine dinucleotide-dependent enzyme FerB from Paracoccus denitrificans reduces a range of sub strates, including chromate, ferric complexes, benzoquinones and naphtoquinones. The reduced form of nicotinamide adenine dinucleotide serves as a source of electrons. Recombinant unmodified and selenometionine-substituted (Se-Met) FerB derivatives were crystallized, diffraction data for both forms were collected and the phase problem for Se-Met FerB dimer was solved by three-wavelength multiple anomalous dispersion, followed by the combination of molecular replacement and single-wavelength anomalous diffraction phasing. A molecular replacement solution of unmodified FerB tetramer was obtained using Se-Met structure as a search model.
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