Crystallization of Pseudomonas aeruginosa AHL synthase LasI using beta-turn crystal engineering.
Crystallization of Pseudomonas aeruginosa AHL synthase LasI using beta-turn crystal engineering.
复制标题
使用 β 转晶体工程对铜绿假单胞菌 AHL 合酶 LasI 进行结晶。
DOI：
10.1107/s0907444903028300
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Churchill,MairEA
中科院分区：
文献类型：
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作者：
Gould,TyA;Watson,WilliamT;Choi,KyoungHee;Schweizer,HerbertP;Churchill,MairEA
在革兰氏阴性菌中，细胞间通讯和毒力调节是由可扩散的化学信号酰基高丝氨酸内酯（阿勒）介导的。阿勒合酶从底物S-腺苷-L-甲硫氨酸和酰基-酰基载体蛋白产生多种AHL。LasI，来自铜绿假单胞菌的阿勒合酶，具有低溶解度，尽管进行了广泛的结晶试验，但未能结晶。基于先前确定的阿勒合酶EsaI的结构，通过将紧转角中的残基重新工程化以产生I′ β-转角来产生活性可溶性LasI。所得的蛋白质是活性的，比野生型LasI更稳定，并且已经在立方空间群F23中结晶，晶胞参数a = B = c = 154.90 Ω。
In Gram-negative bacteria, intercellular communication and virulence regulation is mediated by the diffusible chemical signal acyl-homoserine-l-lactone (AHL). The AHL synthase enzymes produce a variety of AHLs from the substrates S-adenosyl-l-methionine and acyl-acyl carrier protein. LasI, the AHL synthase from Pseudomonas aeruginosa, has low solubility and has failed to crystallize despite extensive crystallization trials. Based on the previously determined structure of the AHL synthase EsaI, active soluble LasI was produced by re-engineering residues in a tight turn to produce a type I′ β-turn. The resulting protein is active, more stable than the wild-type LasI and has been crystallized in the cubic space group F23, with unit-cell parameters a = b = c = 154.90 Å.