铜绿假单胞菌PAO1中响应调节蛋白PA3346的受体和茎域（PA3346RS）的纯化、结晶及初步X射线晶体学分析
Purification, crystallization and preliminary X-ray crystallographic analysis of the receiver and stalk domains (PA3346RS) of the response regulator PA3346 from Pseudomonas aeruginosa PAO1.
作者信息
Wu Pei-Hsiu, Hsu Jye-Jin, Chiang Ting-Wei, Hsieh Yin-Cheng, Chang Hwan-You, Chang Shou-Lin, Chen Chun-Jung
机构信息
Life Science Group, Scientific Research Division, National Synchrotron Radiation Research Center, Hsinchu, Taiwan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Aug 1;67(Pt 8):937-40. doi: 10.1107/S1744309111023931. Epub 2011 Jul 21.
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 P4(1) or P4(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%.
应答调节因子PA3346的调节结构域（PA3346RS）由接收结构域和茎部结构域组成，它需要磷酸化才能在镁离子作为辅因子的情况下被激活，从而调节下游蛋白磷酸酶活性，以调控铜绿假单胞菌PAO1中的群体游动性。总分子量为25.3 kDa的融合标签重组PA3346RS已在大肠杆菌中过表达，通过Ni(2+)-NTA和Q-Sepharose离子交换柱进行纯化，并采用悬滴气相扩散法进行结晶。从PA3346RS晶体收集到分辨率为2.0 Å的X射线衍射数据。该晶体属于空间群P4(1)或P4(3)，晶胞参数a = 82.38，c = 73.34 Å。初步分析表明，不对称单元中存在PA3346RS二聚体，溶剂含量为48.6%。
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