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Artículo

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

Klinke, SebastianIcon ; Foos, Nicolas; Rinaldi, Jimena JulietaIcon ; Paris, GastónIcon ; Goldbaum, Fernando AlbertoIcon ; Legrand, Pierre; Guimarães, Beatriz G; Thompson, Andrew
Fecha de publicación: 07/2015
Editorial: Wiley
Revista: Acta Crystallographica Section D-biological Crystallography
ISSN: 0907-4449
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Biología Celular, Microbiología

Resumen

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.
Palabras clave: Experimental Phasing , Anomalous Signal , Two-Component System , Hwe Family
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/10489
URL: http://scripts.iucr.org/cgi-bin/paper?S1399004715007622
DOI: https://doi.org/10.1107/S1399004715007622
Colecciones
Articulos(IIBBA)
Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)
Citación
Klinke, Sebastian; Foos, Nicolas; Rinaldi, Jimena Julieta; Paris, Gastón; Goldbaum, Fernando Alberto; et al.; 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; Wiley; Acta Crystallographica Section D-biological Crystallography; 71; Pt 7; 7-2015; 1433-1443
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