Extending the eNOE data set of large proteins by evaluation of NOEs with unresolved diagonals
Extending the eNOE data set of large proteins by evaluation of NOEs with unresolved diagonals
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DOI：
10.1007/s10858-015-9917-8
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发表时间：
2015-05-01
影响因子：
2.7
通讯作者：
Voegeli, Beat
中科院分区：
文献类型：
--
作者：
Chi, Celestine N.;Strotz, Dean;Voegeli, Beat
以原子分辨率表示蛋白质的空间采样是理解其功能的基础。NMR已被确立为实现小蛋白质这一目标的最佳技术。然而，可访问的信息内容随着蛋白质大小的增加而迅速恶化。我们最近已经证明，对于小蛋白质的距离限制，精度小于0.1，可以通过用精确的NOE（eNOE）取代传统的半定量核奥弗豪泽效应（NOE）来获得。高质量的数据使我们能够计算由多个而不是单个状态组成的小模型蛋白GB3的结构集合。分析仅限于小蛋白质，因为无法使用具有未分辨对角峰的自旋NOE。在这里，我们提出了一个简单的方法来翻译这样的NOE到正确的上限距离限制，这打开了访问更大的生物分子。我们证明，对于16 kDa亲环素A，这种限制的集合将原始的1254个eNOE扩展到3471个。
The representation of a protein's spatial sampling at atomic resolution is fundamental for understanding its function. NMR has been established as the best-suited technique toward this goal for small proteins. However, the accessible information content rapidly deteriorates with increasing protein size. We have recently demonstrated that for small proteins distance restraints with an accuracy smaller than 0.1 can be obtained by replacing traditional semi-quantitative Nuclear Overhauser Effects (NOEs) with exact NOEs (eNOE). The high quality of the data allowed us to calculate structural ensembles of the small model protein GB3 consisting of multiple rather than a single state. The analysis has been limited to small proteins because NOEs of spins with unresolved diagonal peaks cannot be used. Here we propose a simple approach to translate such NOEs into correct upper distance restraints, which opens access to larger biomolecules. We demonstrate that for 16 kDa cyclophilin A the collection of such restraints extends the original 1254 eNOEs to 3471.