铜绿假单胞菌PAO1生物合成丙氨酸消旋酶的结晶及初步X射线研究
Crystallization and preliminary X-ray study of biosynthetic alanine racemase from Pseudomonas aeruginosa PAO1.
作者信息
Zhou Honggang, Li Zhenzhen, Zhang Guofang, Xu Shujing, Tang Zhaona, Zhu Xianming, Dong Hui, Ju Jiansong
机构信息
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy, Nankai University, Tianjin 300071, People's Republic of China.
Tianjin International Joint Academy of Biomedicine, Tianjin 300457, People's Republic of China.
出版信息
Acta Crystallogr F Struct Biol Commun. 2014 Dec 1;70(Pt 12):1616-9. doi: 10.1107/S2053230X14022432. Epub 2014 Nov 14.
Biosynthetic alanine racemase (AlrPA) from Pseudomonas aeruginosa PAO1 carrying a His6 tag was expressed in Escherichia coli BL21 (DE3) cells and purified by Ni(2+)-chelating affinity and anion-exchange chromatography for X-ray crystallographic analysis. Crystals were grown by the hanging-drop vapour-diffusion method at 289 K in a solution consisting of 4%(v/v) Tacsimate pH 5.0, 14%(w/v) polyethylene glycol 3350 with a protein concentration of 8 mg ml(-1). The crystal diffracted to 2.76 Å resolution and belonged to the orthorhombic space group P212121, with unit-cell parameters a = 74.12, b = 76.97, c = 154.80 Å, α = β = γ = 90°.
携带His6标签的铜绿假单胞菌PAO1的生物合成丙氨酸消旋酶（AlrPA）在大肠杆菌BL21（DE3）细胞中表达，并通过Ni(2+)螯合亲和层析和阴离子交换层析进行纯化，用于X射线晶体学分析。晶体通过悬滴气相扩散法在289 K下于由4%（v/v）pH 5.0的Tacsimate、14%（w/v）聚乙二醇3350组成的溶液中生长，蛋白质浓度为8 mg ml(-1)。该晶体衍射分辨率达到2.76 Å，属于正交晶系空间群P212121，晶胞参数a = 74.12、b = 76.97、c = 154.80 Å，α = β = γ = 90°。
Zotero 插件