Determination of the crystal structure and active residues of FabV, the enoyl-ACP reductase from Xanthomonas oryzae.
Determination of the crystal structure and active residues of FabV, the enoyl-ACP reductase from Xanthomonas oryzae.
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米黄单胞菌烯酰 ACP 还原酶 FabV 晶体结构和活性残基的测定
DOI：
10.1371/journal.pone.0026743
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Jiang T
中科院分区：
文献类型：
--
作者：
Li H;Zhang X;Bi L;He J;Jiang T
研究背景烯酰-ACP还原酶（ENR）催化细菌脂肪酸延伸循环中的最后一个还原反应，是一种很好的抗菌靶标候选物。FabV是最近发现的ENR类，但我们缺乏有关的原子结构和参与还原酶活性的关键残基的信息，除了已知的Y-X8-K活性位点基序中保守的酪氨酸和赖氨酸残基。方法/主要发现在这里，我们报告的晶体结构的FabV从黄单胞菌（xoFabV）。该酶的晶体结构已解析到1.6 μ m分辨率，空间群为P212121。xoFabV的模型由不对称单元中的一个单体组成，该不对称单元由13个α-螺旋和11个β-链组成，代表了典型的Rossmann折叠结构。xoFabV的Y-X8-K活性位点基序和ecFabI的Y-X6-K基序中保守的酪氨酸（Y236）和赖氨酸（K245）残基的位置非常相似。然而，xoFabV中的Y236和ecFabI中的Y156的构象是不同的。基于结构的定点突变和酶活性测定表明，除了Y-X8-K基序中保守的Y236和K245外，Y 53，D111和Y226是与还原酶活性有关的关键残基，F113和T276对酶功能也很重要。此外，在xoFabV中，位于来自鼻疽伯克霍尔德氏菌的FabV（bmFabV）中紧接Y-X8-K基序之后的拟议活性赖氨酸被改变为无活性的V246。结论/意义我们确定了FabV酶的第一晶体结构，并确定了几个残基对其酶活性很重要。这些发现为开发针对霍乱弧菌、伯克霍尔德氏菌和黄单胞菌等病原菌的特异性抗菌抑制剂奠定了坚实的基础。
Background Enoyl-ACP reductase (ENR) catalyses the last reduction reaction in the fatty acid elongation cycle in bacteria and is a good antimicrobial target candidate. FabV is the most recently discovered class of ENR, but we lack information about the atomic structure and the key residues involved in reductase activity except for the known conserved tyrosine and lysine residues in the Y-X8-K active site motif. Methodology/Principal Findings Here we report the crystal structure of FabV from Xanthomonas oryzae (xoFabV). The crystal structure of this enzyme has been solved to 1.6 Å resolution in space group P212121. The model of xoFabV consists of one monomer in the asymmetric unit which is composed of 13 α-helices and 11 β-strands, representing a canonical Rossmann fold architecture. Structural comparison presents that the locations of the conserved tyrosine (Y236) and lysine (K245) residues in the Y-X8-K active site motif of xoFabV and the Y-X6-K motif of ecFabI are notably similar. However, the conformations of Y236 in xoFabV and Y156 in ecFabI are distinct. Structure-based site-directed mutagenesis and enzymatic activity assays reveal that in addition to the conserved Y236 and K245 in the Y-X8-K motif, Y53, D111 and Y226 are key residues implicated in the reductase activity, and F113 and T276 are also important for enzyme function. Moreover, a proposed active lysine located immediately after the Y-X8-K motif in FabV from Burkholderia mallei (bmFabV) is altered to an inactive V246 in xoFabV. Conclusions/Significance We determine the first crystal structure of the FabV enzyme and identify several residues important for its enzymatic activity. These findings lay a solid foundation for the development of specific antibacterial inhibitors of the pathogenic bacteria, such as Vibrio cholerae, Burkholderia species and Xanthomonas species.
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影响因子：
5.8
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1107/s0907444904019158
发表时间：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH