Crystallization of the NADH-oxidizing domain of the Na{sup +}-translocating NADH:ubiquinone oxidoreductase from Vibrio cholerae
- Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057 Zürich (Switzerland)
- School of Engineering and Science, International University Bremen, 28759 Bremen (Germany)
- Swiss Light Source at Paul Scherrer Institut, 5232 Villigen PSI (Switzerland)
- Fachbereich Biologie, Universität Konstanz, Postfach M665, Universitätsstrasse 10, 78457 Konstanz (Germany)
The FAD domain of the NqrF subunit from the Na{sup +}-translocating NADH dehydrogenase from V. cholerae has been purified and crystallized. A complete data set was recorded at 3.1 Å. The Na{sup +}-translocating NADH:quinone oxidoreductase (Na{sup +}-NQR) from pathogenic and marine bacteria is a respiratory complex that couples the exergonic oxidation of NADH by quinone to the transport of Na{sup +} across the membrane. The NqrF subunit oxidizes NADH and transfers the electrons to other redox cofactors in the enzyme. The FAD-containing domain of NqrF has been expressed, purified and crystallized. The purified NqrF FAD domain exhibited high rates of NADH oxidation and contained stoichiometric amounts of the FAD cofactor. Initial crystallization of the flavin domain was achieved by the sitting-drop technique using a Cartesian MicroSys4000 robot. Optimization of the crystallization conditions yielded yellow hexagonal crystals with dimensions of 30 × 30 × 70 µm. The protein mainly crystallizes in long hexagonal needles with a diameter of up to 30 µm. Crystals diffract to 2.8 Å and belong to space group P622, with unit-cell parameters a = b = 145.3, c = 90.2 Å, α = β = 90, γ = 120°.
- OSTI ID:
- 22356273
- Journal Information:
- Acta Crystallographica. Section F, Vol. 62, Issue Pt 2; Other Information: PMCID: PMC2150961; PMID: 16511277; PUBLISHER-ID: vr5056; OAI: oai:pubmedcentral.nih.gov:2150961; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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