Construction of ATP-Switched Allosteric Antioxidant Selenoenzyme
Construction of ATP-Switched Allosteric Antioxidant Selenoenzyme
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ATP转换变构抗氧化硒酶的构建
DOI：
10.1021/acscatal.6b03274
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发表时间：
2017-03-01
期刊：
影响因子：
12.9
通讯作者：
Liu, Junqiu
中科院分区：
文献类型：
--
作者：
Pan, Tiezheng;Liu, Yao;Liu, Junqiu
合理重新设计变构蛋白为开发可切换生物催化剂提供了一种有效的策略。通过将计算设计和蛋白质工程相结合，将谷胱甘肽过氧化物酶(GPX)样活性中心精确地整合到腺苷酸激酶(AKE)的变构结构域中，该活性中心含有催化的硒半胱氨酸(SEC)残基和底物结合的Arg残基。该工程酶不仅具有较高的GPX活性，而且还具有对三磷酸腺苷(ATP)反应的催化活性，这种催化活性受其与三磷酸腺苷(ATP)结合时由开放到封闭的构象变化调节。理论和突变分析表明，静电相互作用和范德华(VDW)相互作用对底物识别的协同作用是导致高活性的主要原因。线粒体氧化损伤实验进一步证明了其在亚细胞水平的抗氧化能力，为体内可控催化提供了潜在的应用。
Rational redesign of allosteric protein offers an efficient strategy to develop switchable biocatalysts. By combining the computational design and protein engineering, a glutathione peroxidase (GPx)-like active center that contains the catalytic selenocysteine (Sec) residue and substrate-binding Arg residue was precisely incorporated into the allosteric domain of adenylate kinase (AKe). The engineered selenoenzyme shows not only high GPx activity but also adenosine triphosphate (ATP)-responsive catalytic property, which is regulated by its opened to closed conformational change upon ATP binding. Theoretical and mutational analysis reveals that the synergistic effect of electrostatic interactions and van der Waals (vdW) interactions for substrate recognition is a major contribution to the high activity. The mitochondrial oxidative damage experiment further demonstrated its antioxidant ability at the subcellular level, offering a potential application toward controllable catalysis in vivo.