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首页> 外文期刊>Nucleic Acids Research >Structure of the uncomplexed DNA repair enzyme endonuclease VIII indicates significant interdomain flexibility.
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Structure of the uncomplexed DNA repair enzyme endonuclease VIII indicates significant interdomain flexibility.

机译：未复合的DNA修复酶核酸内切酶VIII的结构表明明显的域间柔性。

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摘要

Escherichia coli endonuclease VIII (Nei) excises oxidized pyrimidines from DNA. It shares significant sequence homology and similar mechanism with Fpg, a bacterial 8-oxoguanine glycosylase. The structure of a covalent Nei-DNA complex has been recently determined, revealing critical amino acid residues which are important for DNA binding and catalysis. Several Fpg structures have also been reported; however, analysis of structural dynamics of Fpg/Nei family proteins has been hindered by the lack of structures of uncomplexed and DNA-bound enzymes from the same source. We report a 2.8 A resolution structure of free wild-type Nei and two structures of its inactive mutants, Nei-E2A (2.3 A) and Nei-R252A (2.05 A). All three structures are virtually identical, demonstrating that the mutations did not affect the overall conformation of the protein in its free state. The structures show a significant conformational change compared with the Nei structure in its complex with DNA, reflecting a approximately 50 degrees rotation of the two main domains of the enzyme. Such interdomain flexibility has not been reported previously for any DNA glycosylase and may present the first evidence for a global DNA-induced conformational change in this class of enzymes. Several local but functionally relevant structural changes are also evident in other parts of the enzyme.
机译：大肠杆菌核酸内切酶VIII（Nei）从DNA中切除氧化的嘧啶。它与细菌8-氧代鸟嘌呤糖基化酶Fpg具有明显的序列同源性和相似的机制。最近已经确定了共价Nei-DNA复合物的结构，揭示了对于DNA结合和催化很重要的关键氨基酸残基。还报道了几种Fpg结构。然而，Fpg / Nei家族蛋白的结构动力学分析由于缺乏来自同一来源的未复合酶和与DNA结合的酶的结构而受到阻碍。我们报告了一个2.8 A的自由野生型Nei及其非活性突变体的两个结构，Nei-E2A（2.3 A）和Nei-R252A（2.05 A）的解析结构。这三个结构实际上是相同的，这表明突变并不影响处于游离状态的蛋白质的整体构象。与Nei结构与DNA的复合物相比，该结构显示出显着的构象变化，反映出酶的两个主要结构域旋转了大约50度。这种域间的灵活性以前没有针对任何DNA糖基化酶的报道，可能为此类酶中DNA诱导的整体构象变化提供了第一个证据。酶的其他部分也有一些局部但功能相关的结构变化。

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