Crystallization and initial X-ray diffraction studies of the flavoenzyme NAD(P)H:(acceptor) oxidoreductase (FerB) from the soil bacterium Paracoccus denitrificans

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2010-04, Vol.66 (4), p.431-434
• Main Authors: Klumpler, Tomáš, Sedláček, Vojtěch, Marek, Jaromír, Wimmerová, Michaela, Kučera, Igor
• Format: Article
• Language: English
• Subjects: Bacteria; Crystallization; Crystallization Communications; Crystallography, X-Ray; flavoenzymes; NADH Dehydrogenase - chemistry; Paracoccus denitrificans; Paracoccus denitrificans - enzymology; quinone reductases; Soil Microbiology
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309110005099

The flavin‐dependent enzyme FerB from Paracoccus denitrificans reduces a broad range of compounds, including ferric complexes, chromate and most notably quinones, at the expense of the reduced nicotinamide adenine dinucleotide cofactors NADH or NADPH. Recombinant unmodified and SeMet‐substituted FerB were crystallized under similar conditions by the hanging‐drop vapour‐diffusion method with microseeding using PEG 4000 as the precipitant. FerB crystallized in several different crystal forms, some of which diffracted to approximately 1.8 Å resolution. The crystals of native FerB belonged to space group P21, with unit‐cell parameters a = 61.6, b = 110.1, c = 65.2 Å, β = 118.2° and four protein molecules in the asymmetric unit, whilst the SeMet‐substituted form crystallized in space group P21212, with unit‐cell parameters a = 61.2, b = 89.2, c = 71.5 Å and two protein molecules in the asymmetric unit. Structure determination by the three‐wavelength MAD/MRSAD method is now in progress.