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首页> 外文期刊>Acta crystallographica.Section D. Biological crystallography >The 1.8 angstrom resolution structure of hydroxycinnamoyl-coenzyme A hydratase-lyase (HCHL) from Pseudomonas fluorescens, an enzyme that catalyses the transformation of feruloyl-coenzyme A to vanillin
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The 1.8 angstrom resolution structure of hydroxycinnamoyl-coenzyme A hydratase-lyase (HCHL) from Pseudomonas fluorescens, an enzyme that catalyses the transformation of feruloyl-coenzyme A to vanillin

机译：1.8埃分辨率的结构hydroxycinnamoyl-coenzyme一hydratase-lyase(HCHL)的荧光假单胞菌,一种酶催化作用的变换feruloyl-coenzyme,香兰素

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摘要

The crystal structure of hydroxycinnamoyl-CoA hydrataselyase (HCHL) from Pseudomonas fluorescens AN103 has been solved to 1.8 A resolution. HCHL is a member of the crotonase superfamily and catalyses the hydration of the acyl-CoA thioester of ferulic acid [3-(4-hydroxy-3-methoxy-phenyl)prop-2-enoic acid] and the subsequent retro-aldol cleavage of the hydrated intermediate to yield vanillin (4-hydroxy3- methoxy-benzaldehyde). The structure contains 12 molecules in the asymmetric unit, in which HCHL assumes a hexameric structure of two stacked trimers. The substrate, feruloyl-CoA, was modelled into the active site based on the structure of enoyl-CoA hydratase bound to the feruloyl-CoA-like substrate 4-(N,N-dimethylamino)-cinnamoyl-CoA (PDB code 1ey3). Feruloyl-CoA was bound in this model between helix 3 of the A subunit and helix 9 of the B subunit. A highly ordered structural water in the HCHL structure coincided with the thioester carbonyl of feruloyl-CoA in the model, suggesting that the oxyanion hole for stabilization of a thioester-derived enolate, characteristic of coenzyme-A dependent members of the crotonase superfamily, is conserved. The model also suggested that a strong hydrogen bond between the phenolic hydroxyl groups of feruloyl-CoA and BTyr239 may be an important determinant of the enzyme's ability to discriminate between the natural substrate and cinnamoyl-CoA, which is not a substrate.
机译：hydroxycinnamoyl-CoA的晶体结构1.8荧光AN103已经解决了决议。总科和催化作用的水合作用阿魏酸的酰coa硫代酸酯(3 - (4-hydroxy-3-methoxy-phenyl) prop-2-enoic酸)以及随后retro-aldol乳沟的水化中间产生香兰素(4-hydroxy3 - methoxy-benzaldehyde)。包含12个分子不对称单位,HCHL假定两个起点周围形成六聚体结构三。模仿到活动网站的基础上enoyl-CoA酶绑定到的结构(4) - N, N-dimethylamino -cinnamoyl-CoA (PDB代码1 ey3)。螺旋3的一个亚基与螺旋9的B亚基。在正值HCHL结构硫代酸酯羰基feruloyl-CoA的模型,这表明oxyanion洞稳定的thioester-derived烯醇化物,辅酶a的特征相关的成员的crotonase总科,是守恒的。模型还表明,强烈的氢键酚羟基之间的feruloyl-CoA和BTyr239可能是一个重要的行列式的酶的能力自然基质和之间的区别cinnamoyl-CoA,这不是一个衬底。

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