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 S, FOOS N, RINALDI JJ, PARIS G, GOLDBAUM FA, LEGRAND P, GUIMARAES BG AND THOMPSON A

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2015

0907-4449

The histidine kinase (HK) domain belonging to the light?oxygen?voltagehistidine kinase (LOV-HK) from Brucella abortus is a member of the HWEfamily, for which no structural information is available, and has low sequenceidentity (20%) to the closest HK present in the PDB. The ?off-edge? S-SADmethod in macromolecular X-ray crystallography was used to solve the structureof the HK domain from LOV-HK at low resolution from crystals in a lowsymmetryspace group (P21) and with four copies in the asymmetric unit(108 kDa). Data were collected both from multiple crystals (diffraction limitvarying from 2.90 to 3.25 A ˚ ) and from multiple orientations of the same crystal,using the -geometry goniostat on SOLEIL beamline PROXIMA 1, to obtain?true redundancy?. Data from three different crystals were combined forstructure determination. An optimized HK construct bearing a shorter cloningartifact yielded crystals that diffracted X-rays to 2.51 A ˚ resolution and that wereused for final refinement of the model. Moreover, a thorough a posteriorianalysis using several different combinations of data sets allowed us toinvestigate the impact of the data-collection strategy on the success of thestructure determination.