Modulating and evaluating receptor promiscuity through directed evolution and modeling.
Modulating and evaluating receptor promiscuity through directed evolution and modeling.
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通过定向进化和建模来调节和评估受体混杂。
DOI：
10.1093/protein/gzx018
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Tyo,KeithEJ
中科院分区：
文献类型：
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作者：
Stainbrook,SarahC;Yu,JessicaS;Reddick,MichaelP;Bagheri,Neda;Tyo,KeithEJ
G蛋白偶联受体(GPCRs)的混杂在疾病、药理学和生物传感中具有广泛的意义。对于蛋白质工程来说，混杂是一个特别重要的考虑因素，在蛋白质工程中，调节和模拟混杂的能力对于开发理想的蛋白质是必不可少的。在这里，我们介绍了(I)使用定向进化来修正gpr的混杂，以及(Ii)使用定量结构-活性关系模型和分组-穷举特征选择来预测受体反应和识别重要的多肽特征的方法。我们将这些方法应用于酵母信息素受体Ste2及其天然配体α-因子。利用定向进化，我们创建了针对α因子变体文库的特异性改变的Ste2突变体。然后，我们使用疏水、空间和电子性质的矢量和偏最小二乘回归来表征受体-配体的相互作用，确定重要的配体位置和性质，并预测受体对新配体的反应。定向进化和计算分析结合在一起，能够控制和评估GPCR的混杂。这些方法应该广泛用于GPCRs和其他蛋白质-小分子相互作用的研究和工程设计。
The promiscuity of G-protein-coupled receptors (GPCRs) has broad implications in disease, pharmacology and biosensing. Promiscuity is a particularly crucial consideration for protein engineering, where the ability to modulate and model promiscuity is essential for developing desirable proteins. Here, we present methodologies for (i) modifying GPCR promiscuity using directed evolution and (ii) predicting receptor response and identifying important peptide features using quantitative structure-activity relationship models and grouping-exhaustive feature selection. We apply these methodologies to the yeast pheromone receptor Ste2 and its native ligand α-factor. Using directed evolution, we created Ste2 mutants with altered specificity toward a library of α-factor variants. We then used the Vectors of Hydrophobic, Steric, and Electronic properties and partial least squares regression to characterize receptor-ligand interactions, identify important ligand positions and properties, and predict receptor response to novel ligands. Together, directed evolution and computational analysis enable the control and evaluation of GPCR promiscuity. These approaches should be broadly useful for the study and engineering of GPCRs and other protein-small molecule interactions.