title: S-SAD phasing of monoclinic histidine kinase from Brucella abortus combining data from multiple crystals and orientations: an example of data-collection strategy and a posteriori analysis of different data combinations.
abstract: The histidine kinase (HK) domain belonging to the light-oxygen-voltage histidine kinase (LOV-HK) from Brucella abortus is a member of the HWE family, for which no structural information is available, and has low sequence identity (20%) to the closest HK present in the PDB. The `off-edge' S-SAD method in macromolecular X-ray crystallography was used to solve the structure of the HK domain from LOV-HK at low resolution from crystals in a low-symmetry space group (P21) and with four copies in the asymmetric unit (∼108 kDa). Data were collected both from multiple crystals (diffraction limit varying from 2.90 to 3.25 Å) and from multiple orientations of the same crystal, using the κ-geometry goniostat on SOLEIL beamline PROXIMA 1, to obtain `true redundancy'. Data from three different crystals were combined for structure determination. An optimized HK construct bearing a shorter cloning artifact yielded crystals that diffracted X-rays to 2.51 Å resolution and that were used for final refinement of the model. Moreover, a thorough a posteriori analysis using several different combinations of data sets allowed us to investigate the impact of the data-collection strategy on the success of the structure determination.
authors.affiliation: Fundación Instituto Leloir, IIBBA-CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina.
authors.forename: Sebastián
authors.initials: S
authors.lastname: Klinke
authors.affiliation: Experimental Division, Synchrotron SOLEIL, BP 48, L'Orme des Merisiers, 91192 Gif-sur-Yvette, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Foos
authors.affiliation: Fundación Instituto Leloir, IIBBA-CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina.
authors.forename: Jimena J
authors.initials: JJ
authors.lastname: Rinaldi
authors.affiliation: Fundación Instituto Leloir, IIBBA-CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina.
authors.forename: Gastón
authors.initials: G
authors.lastname: Paris
authors.affiliation: Fundación Instituto Leloir, IIBBA-CONICET, Avenida Patricias Argentinas 435, C1405BWE Buenos Aires, Argentina.
authors.forename: Fernando A
authors.initials: FA
authors.lastname: Goldbaum
authors.affiliation: Experimental Division, Synchrotron SOLEIL, BP 48, L'Orme des Merisiers, 91192 Gif-sur-Yvette, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Legrand
authors.affiliation: Experimental Division, Synchrotron SOLEIL, BP 48, L'Orme des Merisiers, 91192 Gif-sur-Yvette, France.
authors.forename: Beatriz G
authors.initials: BG
authors.lastname: Guimarães
authors.affiliation: Experimental Division, Synchrotron SOLEIL, BP 48, L'Orme des Merisiers, 91192 Gif-sur-Yvette, France.
authors.forename: Andrew
authors.initials: A
authors.lastname: Thompson
mesh_terms: D002003:Brucella abortus / Q000737:chemistry / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D000071677:Histidine Kinase; D008958:Models, Molecular; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry*; D017434:Protein Structure, Tertiary
chemical_list: D011494:Protein Kinases; D000071677:Histidine Kinase
keywords: HWE family; anomalous signal; experimental phasing; two-component system