Prediction of Function Determining and Buried Residues Through Analysis of Saturation Mutagenesis Datasets.
Prediction of Function Determining and Buried Residues Through Analysis of Saturation Mutagenesis Datasets.
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DOI：
10.3389/fmolb.2021.635425
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发表时间：
2021
影响因子：
5
通讯作者：
Varadarajan R
中科院分区：
文献类型：
--
作者：
Bhasin M;Varadarajan R
突变扫描可用于探测大量点突变对蛋白质功能的影响。受突变影响的位置主要位于直接参与功能的埋藏或暴露残基，以下称为活性位点残基。在缺乏先验结构信息的情况下，区分这两类残基并不容易。我们策划并分析了一组十二个已发布的深度突变扫描数据集。分析揭示了埋藏和暴露位置的突变敏感性和替代偏好的差异模式。通过结合预测的基于序列的可及性值，可以促进仅根据突变敏感性数据预测埋藏位点。对于活性位点残基，我们观察到平均灵敏度、特异性和准确度分别为 61%、90% 和 88%。对于埋藏的残基，相应的数字分别为 59%、90% 和 84%，而对于暴露的非活性位点残基，相应的数字分别为 98%、44% 和 82%。我们还确定了不遵循这些总体趋势并且可能需要进一步实验重新验证的位置。该分析强调了深度突变扫描即使在没有使用传统结构确定技术确定三维结构的情况下也能提供重要的结构和功能见解的能力，并讨论了该方法的一些局限性。
Mutational scanning can be used to probe effects of large numbers of point mutations on protein function. Positions affected by mutation are primarily at either buried or at exposed residues directly involved in function, hereafter designated as active-site residues. In the absence of prior structural information, it has not been easy to distinguish between these two categories of residues. We curated and analyzed a set of twelve published deep mutational scanning datasets. The analysis revealed differential patterns of mutational sensitivity and substitution preferences at buried and exposed positions. Prediction of buried-sites solely from the mutational sensitivity data was facilitated by incorporating predicted sequence-based accessibility values. For active-site residues we observed mean sensitivity, specificity and accuracy of 61, 90 and 88% respectively. For buried residues the corresponding figures were 59, 90 and 84% while for exposed non active-site residues these were 98, 44 and 82% respectively. We also identified positions which did not follow these general trends and might require further experimental re-validation. This analysis highlights the ability of deep mutational scans to provide important structural and functional insights, even in the absence of three-dimensional structures determined using conventional structure determination techniques, and also discuss some limitations of the methodology.
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影响因子：
3.3
作者：
Gray VE;Hause RJ;Fowler DM
通讯作者：
Fowler DM
影响因子：
48
作者：
Fowler, Douglas M.;Fields, Stanley
通讯作者：
Fields, Stanley
影响因子：
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作者：
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作者：
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通讯作者：
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作者：
Bajaj, Kanika;Dewan, Pooja C.;Varadarajan, Raghavan
通讯作者：
Varadarajan, Raghavan