Open AccessPurification, crystallization and preliminary X‐ray crystallographic analysis of the receiver and stalk domains (PA3346RS) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1
Author(s) -
Wu PeiHsiu,
Hsu JyeJin,
Chiang TingWei,
Hsieh YinCheng,
Chang HwanYou,
Chang ShouLin,
Chen ChunJung
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111023931
Subject(s) - pseudomonas aeruginosa , chemistry , crystallization , stalk , escherichia coli , regulator , dimer , lysis , crystallography , biology , biochemistry , bacteria , gene , organic chemistry , horticulture , genetics
The regulatory domain (PA3346RS), comprising the receiver and stalk domains, of the response regulator PA3346 requires phosphorylation for activation with magnesium ions as cofactors in order to modulate the downstream protein phosphatase activity for the regulation of swarming motility in Pseudomonas aeruginosa PAO1. Fusion‐tagged recombinant PA3346RS of total molecular mass 25.3 kDa has been overexpressed in Escherichia coli , purified using Ni 2+ –NTA and Q‐Sepharose ion‐exchange columns and crystallized using the hanging‐drop vapour‐diffusion method. X‐ray diffraction data were collected from PA3346RS crystals to 2.0 Å resolution. The crystal belonged to space group P 4 1 or P 4 3 , with unit‐cell parameters a = 82.38, c = 73.34 Å. Preliminary analysis indicated the presence of a dimer of PA3346RS in the asymmetric unit, with a solvent content of 48.6%.