Evolution of a designed retro-aldolase leads to complete active site remodeling.
Evolution of a designed retro-aldolase leads to complete active site remodeling.
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DOI：
10.1038/nchembio.1276
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发表时间：
2013-08
影响因子：
14.8
通讯作者：
Hilvert D
中科院分区：
文献类型：
--
作者：
Giger L;Caner S;Obexer R;Kast P;Baker D;Ban N;Hilvert D
进化的进步往往是由意想不到的创新推动的。计算设计的酶的定向进化表明，剧烈的分子变化也可以驱动原始蛋白质活性位点的优化。通过随机诱变和筛选，人工反转录醛缩酶的比活性提高了4400倍，达到了接近天然酶的催化效率。然而，结构和机理研究表明，由活性赖氨酸和有序水分子组成的工程催化装置意外地被放弃，取而代之的是在优化过程中产生的底物结合口袋中的新赖氨酸残基。最初的硅设计结构，一个机械上混杂的中间体，以及一个最进化的变体，突出了环迁移率和支持官能团在新催化中心出现中的重要性。这种不同反应位点之间的内部竞争可能是许多天然酶早期进化的特征。
Evolutionary advances are often fueled by unanticipated innovation. Directed evolution of a computationally designed enzyme suggests that dramatic molecular changes can also drive the optimization of primitive protein active sites. The specific activity of an artificial retro-aldolase was boosted >4,400 fold by random mutagenesis and screening, affording catalytic efficiencies approaching those of natural enzymes. However, structural and mechanistic studies reveal that the engineered catalytic apparatus, consisting of a reactive lysine and an ordered water molecule, was unexpectedly abandoned in favor of a new lysine residue in a substrate binding pocket created during the optimization process. Structures of the initial in silico design, a mechanistically promiscuous intermediate, and one of the most evolved variants highlight the importance of loop mobility and supporting functional groups in the emergence of the new catalytic center. Such internal competition between alternative reactive sites may have characterized the early evolution of many natural enzymes.
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影响因子：
3.5
作者：
Esposito, G;Vitagliano, L;Salvatore, F
通讯作者：
Salvatore, F
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
56.9
作者：
Heine, A;DeSantis, G;Wilson, IA
通讯作者：
Wilson, IA
影响因子：
3.7
作者：
Richter F;Leaver-Fay A;Khare SD;Bjelic S;Baker D
通讯作者：
Baker D
影响因子：
8
作者：
Althoff, Eric A.;Wang, Ling;Jiang, Lin;Giger, Lars;Lassila, Jonathan K.;Wang, Zhizhi;Smith, Matthew;Hari, Sanjay;Kast, Peter;Herschlag, Daniel;Hilvert, Donald;Baker, David
通讯作者：
Baker, David