A highly stable protein chimera built from fragments of different folds.
A highly stable protein chimera built from fragments of different folds.
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DOI：
10.1093/protein/gzs074
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发表时间：
2012-11
期刊：
影响因子：
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通讯作者：
S. Shanmugaratnam;Simone Eisenbeis;B. Höcker
中科院分区：
文献类型：
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作者：
S. Shanmugaratnam;Simone Eisenbeis;B. Höcker
在进化过程中，蛋白质的复杂性增加了。结构域和亚结构域大小的片段被招募并调整以形成新的蛋白质和新的折叠。这一概念可以用于工程中构建新的蛋白质。我们之前报道了两个古老的蛋白质折叠的片段的组合，一个是黄毒素样蛋白，另一个是(βα)₈-Barrel)蛋白。在这里，我们报告了两次进一步的尝试，从这些折叠的片段中设计出嵌合蛋白。虽然其中一个结构显示出高度聚集的倾向，但另一个结构被证明是高度稳定的、结构良好的蛋白质。在对热和化学变性的稳定性方面，该嵌合体被命名为NarLHisF，优于先前提出的CheYHisF。这是由两个不同的蛋白质折叠构建的嵌合体的第二个实例，它表明重组可以多么容易地导致新蛋白质的发展和多样化--这一机制很可能在进化过程中经常发生。基于失败和成功嵌合体的结果，我们讨论了折叠嵌合体一般设计策略的重要考虑因素。
Proteins increased in complexity during the course of evolution. Domains as well as subdomain-sized fragments were recruited and adapted to form new proteins and novel folds. This concept can be used in engineering to construct new proteins. We previously reported the combination of fragments from two ancient protein folds, a flavodoxin-like and a (βα)₈-barrel protein. Here we report two further attempts at engineering a chimeric protein from fragments of these folds. While one of the constructs showed a high tendency to aggregate, the other turned out to be a highly stable, well-structured protein. In terms of stability against heat and chemical denaturation this chimera, named NarLHisF, is superior to the earlier presented CheYHisF. This is the second instance of a chimera build from two different protein folds, which demonstrates how easily recombination can lead to the development and diversification of new proteins--a mechanism that most likely occurred frequently in the course of evolution. Based on the results of the failed and the successful chimera, we discuss important considerations for a general design strategy for fold chimeras.