Crystallization and preliminary X-ray characterization of the full-length bacteriophytochrome from the plant pathogen Xanthomonas campestris pv. campestris

SEBASTIÁN KLINKE; LISANDRO OTERO; JIMENA RINALDI; SANTIAGO SOSA; BEATRIZ G. GUIMARÃES; WILLIAM E. SHEPARD; FERNANDO A. GOLDBAUM; HERNÁN R. BONOMI

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2014 vol. 70 p. 1636 - 1639

1744-3091

Phytochromes give rise to the largest photosensor family known to date. However, they are underrepresented in the Protein Data Bank. Plant, cyanobacterial, fungal and bacterial phytochromes share a canonical architecture consisting of an N-terminal photosensory module (PAS2-GAF-PHY domains) and a C-terminal variable output module. The bacterium Xanthomonas campestris pv. campestris, a worldwide agricultural pathogen, codes for a single bacteriophytochrome (XccBphP) that holds this canonical architecture, bearing a C-terminal PAS9 domain as the output module. Full-length XccBphP was cloned, expressed and purified to homogeneity by nickel-NTA affinity and size exclusion chromatography and then crystallized at room temperature bound to its cofactor biliverdin. A complete native X-ray diffraction datasetwas collected to a maximum resolution of 3.25 Å. Crystals belong to the space group P43212 with unit-cell parameters a = b = 103.94, c = 344.57 Å, and a dimer in the asymmetric unit. Refinement is underway after solving the structure by molecular replacement.