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首页> 外文期刊>Acta crystallographica, Section F. Structural biology and crystallization communications >Structure of 2-keto-3-deoxy-D-manno-octulosonate-8-phosphate synthase from Pseudomonas aeruginosa
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Structure of 2-keto-3-deoxy-D-manno-octulosonate-8-phosphate synthase from Pseudomonas aeruginosa

机译：铜绿假单胞菌的2-酮-3-脱氧-D-甘露聚糖-八磺酸盐-8-磷酸合酶的结构

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摘要

Pseudomonas aeruginosa is a major cause of opportunistic infection and is resistant to most antibiotics. As part of efforts to generate much-needed new antibiotics, structural studies of enzymes that are critical for the virulence of P. aeruginosa but are absent in mammals have been initiated. 2-Keto-3-deoxy-D-manno-octulosonate-8-phosphate synthase (KDO8Ps), also known as 2-dehydro-3-deoxyphosphooctonate aldolase, is vital for the survival and virulence of P. aeruginosa. This enzyme catalyzes a key step in the synthesis of the lipopolysaccharide (LPS) of most Gram-negative bacteria: the condensation reaction between phosphoenolpyruvate (PEP) and arabinose 5-phosphate to produce 2-keto-3-deoxy-D-manno-octulosonate-8-phosphate (KDO8P). This step is vital for the proper synthesis and assembly of LPS and the survival of P. aeruginosa. Here, the recombinant expression, purification and crystal structure of KDO8Ps from P. aeruginosa are presented. Orthorhombic crystals were obtained by vapor diffusion in sitting drops in the presence of 1 mM phosphoenlpyruvate. The structure reveals the prototypical alpha/beta TIM-barrel structure expected from this family of enzymes and contains a tetramer in the asymmetric unit.
机译：铜绿假单胞菌是机会感染的主要原因，并且对大多数抗生素具有抗性。作为产生急需的新抗生素的努力的一部分，已经开始了对铜绿假单胞菌的毒性至关重要但在哺乳动物中不存在的酶的结构研究。 2-Keto-3-deoxy-D-manno-octulosonate-8-phosphate synthase（KDO8Ps），也称为2-dehydro-3-deoxyphosphooctonate aldolase，对铜绿假单胞菌的存活和致病性至关重要。该酶催化大多数革兰氏阴性细菌的脂多糖（LPS）合成中的关键步骤：磷酸烯醇式丙酮酸（PEP）与阿拉伯糖5-磷酸之间的缩合反应生成2-酮基-3-脱氧-D-甘露聚糖-八辛酸酯-8-磷酸盐（KDO8P）。此步骤对于LPS的正确合成和组装以及铜绿假单胞菌的生存至关重要。在此，展示了来自铜绿假单胞菌的KDO8P的重组表达，纯化和晶体结构。通过在1 mM磷酸烯丙酮酯存在下静坐滴中的蒸汽扩散获得正交晶体。该结构揭示了该酶家族所期望的原型α/βTIM-桶结构，并且在不对称单元中包含四聚体。

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