Predicting repeat protein folding kinetics from an experimentally determined folding energy landscape.
Predicting repeat protein folding kinetics from an experimentally determined folding energy landscape.
复制标题
从实验确定的折叠能量景观预测重复蛋白质折叠动力学。
DOI：
10.1002/pro.9
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Barrick,Doug
中科院分区：
文献类型：
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作者：
Street,TimothyO;Barrick,Doug
Notch锚蛋白结构域是一种重复蛋白，其折叠已通过平衡和动力学测量表征。在先前的工作中，使用截短构建体的平衡折叠自由能来产生实验确定的折叠能量景观（Mello和Barrick，Proc Natl Acad Sci USA 2004;101：14102-14107）。在这里，这个折叠能量景观是用来参数化的动力学模型，其中局部之间的过渡概率部分折叠状态的能量值的基础上的景观。基于景观的模型正确预测了Notch锚蛋白结构域和序列变体的高度多样的实验确定的折叠动力学。这些预测包括双相折叠和双相解折叠、人字形图解折叠分支的曲率、瞬时解折叠中间体的数量、跨越三个数量级的19种变体的相对折叠速率以及由C末端稳定化引起的折叠途径的变化。这些发现表明Notch锚蛋白结构域的折叠途径是选择性的：动力学行为的主要决定因素可以简单地从单个重复序列的局部稳定性推导出来。
The Notch ankyrin domain is a repeat protein whose folding has been characterized through equilibrium and kinetic measurements. In previous work, equilibrium folding free energies of truncated constructs were used to generate an experimentally determined folding energy landscape (Mello and Barrick, Proc Natl Acad Sci USA 2004;101:14102–14107). Here, this folding energy landscape is used to parameterize a kinetic model in which local transition probabilities between partly folded states are based on energy values from the landscape. The landscape‐based model correctly predicts highly diverse experimentally determined folding kinetics of the Notch ankyrin domain and sequence variants. These predictions include monophasic folding and biphasic unfolding, curvature in the unfolding limb of the chevron plot, population of a transient unfolding intermediate, relative folding rates of 19 variants spanning three orders of magnitude, and a change in the folding pathway that results from C‐terminal stabilization. These findings indicate that the folding pathway(s) of the Notch ankyrin domain are thermodynamically selected: the primary determinants of kinetic behavior can be simply deduced from the local stability of individual repeats.