Biophysical Inference of Epistasis and the Effects of Mutations on Protein Stability and Function
Biophysical Inference of Epistasis and the Effects of Mutations on Protein Stability and Function
复制标题
上位性的生物物理推断以及突变对蛋白质稳定性和功能的影响
DOI：
--
复制
发表时间：
2018
影响因子：
10.7
通讯作者：
Jakub Otwinowski
中科院分区：
文献类型：
--
作者：
Jakub Otwinowski
了解蛋白质序列、功能和稳定性之间的关系是生物学中的一个基本问题。许多折叠成特定结构的蛋白质的基本功能是它们与配体结合的能力，可以对数千种突变变体进行检测。然而，结合分析并不区分突变是影响结合界面的稳定性还是影响整体折叠。在这里，我们介绍了一种统计方法，通过结合来自许多突变变体的信息来推断蛋白质如何折叠并与配体结合的详细能量图景。我们将描述结合、未结合和未折叠状态的热力学模型与高质量的蛋白G结构域B1与Ig G-Fc结合的数据进行了拟合。我们推断每个突变的不同折叠和结合能，提供突变如何影响整个蛋白质的结合和稳定性的详细视图。我们准确地推断出每个变体在物理单元中的折叠能量，并通过独立的数据进行验证，而以前的高通量方法只能测量稳定性的间接变化。虽然我们假设了一个相加的序列-能量关系，但由于其与能量的非线性关系，结合分数是上位的。尽管在能量上没有上位性，但我们的模型解释了大部分观察到的结合部分的上位性，其余的上位性识别构象动态区域。
Understanding the relationship between protein sequence, function, and stability is a fundamental problem in biology. The essential function of many proteins that fold into a specific structure is their ability to bind to a ligand, which can be assayed for thousands of mutated variants. However, binding assays do not distinguish whether mutations affect the stability of the binding interface or the overall fold. Here, we introduce a statistical method to infer a detailed energy landscape of how a protein folds and binds to a ligand by combining information from many mutated variants. We fit a thermodynamic model describing the bound, unbound, and unfolded states to high quality data of protein G domain B1 binding to IgG-Fc. We infer distinct folding and binding energies for each mutation providing a detailed view of how mutations affect binding and stability across the protein. We accurately infer the folding energy of each variant in physical units, validated by independent data, whereas previous high-throughput methods could only measure indirect changes in stability. While we assume an additive sequence-energy relationship, the binding fraction is epistatic due its nonlinear relation to energy. Despite having no epistasis in energy, our model explains much of the observed epistasis in binding fraction, with the remaining epistasis identifying conformationally dynamic regions.
登录
查看更多内容
影响因子：
6.8
作者：
Bershtein S;Serohijos AW;Shakhnovich EI
通讯作者：
Shakhnovich EI
DOI：
10.1073/pnas.0406744102
发表时间：
2005-01-18
影响因子：
11.1
作者：
Bloom, JD;Silberg, JJ;Arnold, FH
通讯作者：
Arnold, FH
影响因子：
6.8
作者：
Wright, Peter E.;Dyson, H. Jane
通讯作者：
Dyson, H. Jane
用于蛋白质工程和设计的深度测序方法。
影响因子：
6.8
作者：
Wrenbeck,EmilyE;Faber,MatthewS;Whitehead,TimothyA
通讯作者：
Whitehead,TimothyA
影响因子：
7.8
作者：
Magliery, Thomas J.;Lavinder, Jason J.;Sullivan, Brandon J.
通讯作者：
Sullivan, Brandon J.