Purification, crystallization and preliminary X-ray crystallographic analysis of the phosphatase domain (PA3346PD) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1.

ABSTRACT: The phosphatase domain (PA3346PD) of the response regulator PA3346 modulates the downstream anti-anti-? factor PA3347 to regulate swarming motility in Pseudomonas aeruginosa PAO1. PA3346PD, which comprises the protein phosphatase 2C domain (PP2C), is classified as a Ser/Thr phosphatase of the Mg(2+)- or Mn(2+)-dependent protein phosphatase (PPM) family. The recombinant PA3346PD, with molecular mass 26?kDa, was overexpressed in Escherichia coli, purified on an Ni(2+)-NTA agarose column and crystallized by the sitting-drop vapour-diffusion method. X-ray diffraction data were collected from PA3346PD crystals to a resolution of 2.58?Å and the crystals belonged to space group I4?32 or I4?32, with unit-cell parameter a = 157.61?Å. Preliminary analysis indicates the presence of a monomer of PA3346PD in the asymmetric unit with a solvent content of 58.4%.

SUBMITTER: Chen LY 

PROVIDER: S-EPMC4388179 | biostudies-literature | 2015 Apr

REPOSITORIES: biostudies-literature