Expression, purification, crystallization and preliminary X-ray diffraction analysis of a type II NADH:quinone oxidoreductase from the human pathogen Staphylococcus aureus

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 71; no. 4; pp. 477 - 482
• Main Authors: Rosário, Ana Lúcia, Sena, Filipa V., Batista, Ana P., Oliveira, Tânia F., Athayde, Diogo, Pereira, Manuela M., Brito, José A., Archer, Margarida
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.04.2015; Wiley Subscription Services, Inc
• Subjects: Amino Acid Sequence; Crystallization; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Humans; Molecular Sequence Data; Multienzyme Complexes - biosynthesis; Multienzyme Complexes - chemistry; Multienzyme Complexes - isolation & purification; NADH dehydrogenase; NADH, NADPH Oxidoreductases - biosynthesis; NADH, NADPH Oxidoreductases - chemistry; NADH, NADPH Oxidoreductases - isolation & purification; NDH-II; Research Communications; respiratory chain; Staphylococcus aureus; Staphylococcus aureus - enzymology; X-Ray Diffraction; NADH dehydrogenase; NDH-II; Staphylococcus aureus; respiratory chain
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X15005178

In recent years, type II NADH dehydrogenases (NDH‐IIs) have emerged as potential drug targets for a wide range of human disease causative agents. In this work, the NDH‐II enzyme from the Gram‐positive human pathogen Staphylococcus aureus was recombinantly expressed in Escherichia coli, purified, crystallized and a crystallographic data set was collected at a wavelength of 0.873 Å. The crystals belonged to the orthorhombic space group P212121, with unit‐cell parameters a = 81.8, b = 86.0, c = 269.9 Å, contained four monomers per asymmetric unit and diffracted to a resolution of 3.32 Å. A molecular‐replacement solution was obtained and model building and refinement are currently under way.