Recurring sequence-structure motifs in (βα)8-barrel proteins and experimental optimization of a chimeric protein designed based on such motifs
Recurring sequence-structure motifs in (βα)8-barrel proteins and experimental optimization of a chimeric protein designed based on such motifs
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(β α)(8)-桶蛋白中重复的序列结构基序以及基于此类基序设计的嵌合蛋白的实验优化
DOI：
10.1016/j.bbapap.2016.11.001
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发表时间：
2017-02-01
影响因子：
3.2
通讯作者：
Liu，Haiyan
中科院分区：
文献类型：
--
作者：
Wang，Jichao;Zhang，Tongchuan;Liu，Haiyan
产生新的人工蛋白质的一种有趣的方式是将来自天然蛋白质的序列基序联合收割机组合，模拟由包含重复基序的天然蛋白质所暗示的进化路径。采用基于结构分析的序列聚类方法对TIM桶蛋白的βα和αβ模块进行了分析。确定了一些优选的基序。通过使用重复出现的元素作为相互兼容的接口，设计了嵌合TIM。设计的TIM蛋白的折叠性，然后显着提高了六轮的定向进化。熔融温度提高了20 ℃以上。各种特征表明，所得蛋白质是良好折叠的。我们的分析提供了一个肽基序库，这对不同的蛋白质工程研究是潜在有用的。利用重复模体作为界面连接部分天然蛋白质的蛋白质工程策略可以应用于其他蛋白质折叠。
An interesting way of generating novel artificial proteins is to combine sequence motifs from natural proteins, mimicking the evolutionary path suggested by natural proteins comprising recurring motifs. We analyzed the βα and αβ modules of TIM barrel proteins by structure alignment-based sequence clustering. A number of preferred motifs were identified. A chimeric TIM was designed by using recurring elements as mutually compatible interfaces. The foldability of the designed TIM protein was then significantly improved by six rounds of directed evolution. The melting temperature has been improved by more than 20 °C. A variety of characteristics suggested that the resulting protein is well-folded. Our analysis provided a library of peptide motifs that is potentially useful for different protein engineering studies. The protein engineering strategy of using recurring motifs as interfaces to connect partial natural proteins may be applied to other protein folds.