Increased Diels-Alderase activity through backbone remodeling guided by Foldit players.
Increased Diels-Alderase activity through backbone remodeling guided by Foldit players.
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DOI：
10.1038/nbt.2109
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发表时间：
2012-01-22
影响因子：
46.9
通讯作者：
中科院分区：
文献类型：
--
作者：
计算机酶设计为可再生燃料、药物和化学品的生产带来了希望。从头设计的酶已经为几个反应产生了催化剂，但催化效率低于自然产生的酶1-4。在这里，我们报告了使用众包来通过对结构的功能重塑来提高通过计算设计的酶的活性。在线游戏Foldit 5，6的玩家被要求重新塑造通过计算设计的双分子Diels-Alderase 3的主干，以实现与底物的额外相互作用。几次重复的设计和鉴定产生了一个24个残基的螺旋-转弯-螺旋基序，包括13个残基的插入，使酶活性提高了18倍以上。X射线结晶学分析表明，大插入物采用Foldit模型中的螺旋-转角-螺旋结构。这些结果表明，人类对设计问题的创造力可以从日常生活的宏观问题延伸到不太熟悉的分子尺度蛋白质设计问题。
Computational enzyme design holds promise for the production of renewable fuels, drugs, and chemicals. De novo enzyme design has generated catalysts for several reactions, but with lower catalytic efficiencies than naturally occurring enzymes 1–4. Here we report the use of crowdsourcing to enhance the activity of a computationally designed enzyme through the functional remodeling of its structure. Players of the online game Foldit 5, 6 were challenged to remodel the backbone of a computationally designed bimolecular Diels-Alderase 3 to enable additional interactions with substrates. Several iterations of design and characterization generated a 24 residue helix-turn-helix motif, including a 13 residue insertion, that increased enzyme activity over 18-fold. X-ray crystallography showed that the large insertion adopts a helix-turn-helix structure positioned as in the Foldit model. These results demonstrate that human creativity with design problems can extend beyond the macroscopic problems of everyday life to less familiar molecular scale protein design problems.
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影响因子：
64.8
作者：
通讯作者：
--
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
DOI：
10.1073/pnas.0703434104
发表时间：
2007-05-29
影响因子：
11.1
作者：
Religa, Tomasz L.;Johnson, Christopher M.;Fersht, Alan R.
通讯作者：
Fersht, Alan R.
影响因子：
2.9
作者：
Bar-Even, Arren;Noor, Elad;Milo, Ron
通讯作者：
Milo, Ron
影响因子：
56.9
作者：
Park, HS;Nam, SH;Kim, HS
通讯作者：
Kim, HS