title: 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-σ 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 I4₁32 or I4₃32, 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%.
authors: Institute of Biotechnology, National Cheng-Kung University, Tainan 701, Taiwan.
authors: Li Ying
authors: LY
authors: Chen
authors: Life Science Group, Scientific Research Division, National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan.
authors: Pei Hsun
authors: PH
authors: Wu
authors: Life Science Group, Scientific Research Division, National Synchrotron Radiation Research Center, Hsinchu 30076, Taiwan.
authors: Hong Hsiang
authors: HH
authors: Guan
authors: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia.
authors: Hoong Kun
authors: HK
authors: Fun
authors: Institute of Molecular Medicine, National Tsing-Hua University, Hsinchu 30013, Taiwan.
authors: Hwan You
authors: HY
authors: Chang
authors: Institute of Biotechnology, National Cheng-Kung University, Tainan 701, Taiwan.
authors: Chun Jung
authors: CJ
authors: Chen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D010744:Phosphoric Monoester Hydrolases
keywords: Pseudomonas aeruginosa; phosphatase domain; response regulator