Analysis of Large-Scale Mutagenesis Data To Assess the Impact of Single Amino Acid Substitutions.
Analysis of Large-Scale Mutagenesis Data To Assess the Impact of Single Amino Acid Substitutions.
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DOI：
10.1534/genetics.117.300064
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发表时间：
2017-09
期刊：
影响因子：
3.3
通讯作者：
Fowler DM
中科院分区：
文献类型：
--
作者：
Gray VE;Hause RJ;Fowler DM
诱变是一种广泛使用的方法，用于识别对功能或配体结合重要的蛋白质位置。高通量DNA测序和诱变技术的进步已经能够测量许多蛋白质中几乎所有可能的氨基酸取代的影响。由此产生的大规模诱变数据集提供了一个独特的机会，可以得出关于不同氨基酸取代作用的一般结论。因此，我们分析了14种蛋白质的34,373个突变，这些突变的影响是通过大规模诱变方法测量的。蛋氨酸是最耐受性的替代，脯氨酸是最不耐受性的替代。我们发现，一些取代，包括组氨酸和天冬酰胺，最好地再现了其他取代的效果，即使考虑到野生型氨基酸的身份。在不同的结构背景下，组氨酸和天冬酰胺取代的效果也与其他取代的效果最相关。此外，高破坏性取代如天冬氨酸和谷氨酸在检测配体界面位置方面具有最大的区别能力。我们的工作强调了大规模突变数据的实用性，我们的结论可以帮助指导未来的单替代突变扫描。
Mutagenesis is a widely used method for identifying protein positions that are important for function or ligand binding. Advances in high-throughput DNA sequencing and mutagenesis techniques have enabled measurement of the effects of nearly all possible amino acid substitutions in many proteins. The resulting large-scale mutagenesis data sets offer a unique opportunity to draw general conclusions about the effects of different amino acid substitutions. Thus, we analyzed 34,373 mutations in 14 proteins whose effects were measured using large-scale mutagenesis approaches. Methionine was the most tolerated substitution, while proline was the least tolerated. We found that several substitutions, including histidine and asparagine, best recapitulated the effects of other substitutions, even when the identity of the wild-type amino acid was considered. The effects of histidine and asparagine substitutions also correlated best with the effects of other substitutions in different structural contexts. Furthermore, highly disruptive substitutions like aspartic and glutamic acid had the most discriminatory power for detecting ligand interface positions. Our work highlights the utility of large-scale mutagenesis data, and our conclusions can help guide future single substitution mutational scans.