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

KLINKE S, OTERO LH, RINALDI S, SOSA S, GUIMARÃES BG, SHEPARD WE, GOLDBAUM FA AND BONOMI HR

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 architectureconsisting of an N-terminal photosensory module (PAS2?GAF?PHYdomains) and a C-terminal variable output module. The bacterium Xanthomonascampestris pv. campestris, a worldwide agricultural pathogen, codes for asingle bacteriophytochrome (XccBphP) that has this canonical architecture,bearing a C-terminal PAS9 domain as the output module. Full-length XccBphPwas cloned, expressed and purified to homogeneity by nickel?NTA affinity andsize-exclusion chromatography and was then crystallized at room temperaturebound to its cofactor biliverdin. A complete native X-ray diffraction data set wascollected to a maximum resolution of 3.25 A ° . The crystals belonged to spacegroup P43212, with unit-cell parameters a = b = 103.94, c = 344.57 A ° and a dimerin the asymmetric unit. Refinement is underway after solving the structure bymolecular replacement.