Rational Design of Adenylate Kinase Thermostability through Coevolution and Sequence Divergence Analysis.
Rational Design of Adenylate Kinase Thermostability through Coevolution and Sequence Divergence Analysis.
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通过协同进化和序列分歧分析合理设计腺苷酸激酶热稳定性
DOI：
10.3390/ijms22052768
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发表时间：
2021-03-09
影响因子：
5.6
通讯作者：
Tan YW
中科院分区：
文献类型：
--
作者：
Chang J;Zhang C;Cheng H;Tan YW
蛋白质工程在工业和实验室环境中被积极地追求高热稳定性。在众多的蛋白质工程方法中，生物信息学的合理设计提供了理论指导，而无需耗时的实验筛选。然而，大多数合理的设计方法要么依赖于蛋白质三级结构信息，要么精度有限。我们提出了一种基于一级序列的算法来增加蛋白质的耐热性，同时保持其功能。该方法以腺苷酸激酶（ADK）家族为模型体系，鉴定了一系列与热稳定性密切相关的氨基酸位点。基于该方法构建的单点和双点突变体将中温大肠杆菌（E. coli） ADK的热变性温度分别提高了5.5°C和8.3°C，同时在室温下保留了大部分催化功能。此外，构建的突变体在较高温度下具有改善的酶活性。
Protein engineering is actively pursued in industrial and laboratory settings for high thermostability. Among the many protein engineering methods, rational design by bioinformatics provides theoretical guidance without time-consuming experimental screenings. However, most rational design methods either rely on protein tertiary structure information or have limited accuracies. We proposed a primary-sequence-based algorithm for increasing the heat resistance of a protein while maintaining its functions. Using adenylate kinase (ADK) family as a model system, this method identified a series of amino acid sites closely related to thermostability. Single- and double-point mutants constructed based on this method increase the thermal denaturation temperature of the mesophilic Escherichia coli (E. coli) ADK by 5.5 and 8.3 °C, respectively, while preserving most of the catalytic function at ambient temperatures. Additionally, the constructed mutants have improved enzymatic activity at higher temperature.
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影响因子：
6.8
作者：
Magliery TJ
通讯作者：
Magliery TJ
影响因子：
5.6
作者：
Guerois, R;Nielsen, JE;Serrano, L
通讯作者：
Serrano, L
DOI：
10.1107/s2053230x15024619
发表时间：
2016-02
期刊：
Acta crystallographica. Section F, Structural biology communications
影响因子：
--
作者：
Deller MC;Kong L;Rupp B
通讯作者：
Rupp B
影响因子：
5.8
作者：
Huang, Liang-Tsung;Gromiha, M. Michael;Ho, Shinn-Ying
通讯作者：
Ho, Shinn-Ying
影响因子：
5.8
作者：
Pucci, Fabrizio;Bernaerts, Katrien, V;Rooman, Marianne
通讯作者：
Rooman, Marianne