铜绿假单胞菌中一种吩嗪修饰酶——依赖黄素腺嘌呤二核苷酸（FAD）的单加氧酶PhzS的纯化、结晶及初步结构表征
The purification, crystallization and preliminary structural characterization of FAD-dependent monooxygenase PhzS, a phenazine-modifying enzyme from Pseudomonas aeruginosa.
作者信息
Gohain Neelakshi, Thomashow Linda S, Mavrodi Dmitri V, Blankenfeldt Wulf
机构信息
Max-Planck-Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Oct 1;62(Pt 10):989-92. doi: 10.1107/S1744309106034464. Epub 2006 Sep 30.
The blue chloroform-soluble bacterial metabolite pyocyanin (1-hydroxy-5-methyl-phenazine) contributes to the survival and virulence of Pseudomonas aeruginosa, an important Gram-negative opportunistic pathogen of humans and animals. Little is known about the two enzymes, designated PhzM and PhzS, that function in the synthesis of pyocyanin from phenazine-1-carboxylic acid. In this study, the FAD-dependent monooxygenase PhzS was purified and crystallized from lithium sulfate/ammonium sulfate/sodium citrate pH 5.5. Native crystals belong to space group C2, with unit-cell parameters a = 144.2, b = 96.2, c = 71.7 A, alpha = gamma = 90, beta = 110.5 degrees. They contain two monomers of PhzS in the asymmetric unit and diffract to a resolution of 2.4 A. Seleno-L-methionine-labelled PhzS also crystallizes in space group C2, but the unit-cell parameters change to a = 70.6, b = 76.2, c = 80.2 A, alpha = gamma = 90, beta = 110.5 degrees and the diffraction limit is 2.7 A.
蓝色的可溶于氯仿的细菌代谢产物绿脓菌素（1-羟基-5-甲基吩嗪）有助于铜绿假单胞菌的存活和致病性，铜绿假单胞菌是人和动物重要的革兰氏阴性机会致病菌。对于从吩嗪-1-羧酸合成绿脓菌素过程中发挥作用的两种酶，即PhzM和PhzS，人们了解甚少。在本研究中，依赖黄素腺嘌呤二核苷酸（FAD）的单加氧酶PhzS在硫酸锂/硫酸铵/柠檬酸钠pH 5.5条件下被纯化并结晶。天然晶体属于空间群C2，晶胞参数为a = 144.2，b = 96.2，c = 71.7 Å，α = γ = 90°，β = 110.5°。它们在不对称单元中包含两个PhzS单体，衍射分辨率为2.4 Å。硒代-L-甲硫氨酸标记的PhzS也在空间群C2中结晶，但晶胞参数变为a = 70.6，b = 76.2，c = 80.2 Å，α = γ = 90°，β = 110.5°，衍射极限为2.7 Å。
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