Automatic structure-based NMR methyl resonance assignment in large proteins
Automatic structure-based NMR methyl resonance assignment in large proteins
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DOI：
10.1038/s41467-019-12837-8
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发表时间：
2019-10-29
影响因子：
16.6
通讯作者：
Guentert, Peter
中科院分区：
文献类型：
--
作者：
Pritisanac, Iva;Wuerz, Julia M.;Guentert, Peter
同位素标记的甲基基团在大的，否则氘代蛋白质中提供NMR探针。然而，共振归属构成了甲基基NMR更广泛适用性的瓶颈。在这里，我们提出了自动化的甲基FLYA方法的甲基基团的分配是基于甲基-甲基核奥弗豪泽效应光谱（NOESY）的峰列表。将MethylFLYA应用于总共包含708个同位素标记的甲基的五种蛋白质（28-358 kDa），其中612个贡献NOESY交叉峰。MethylFLYA确定了488个甲基，即80%的NOESY数据。其中，459个同意参考文献，6个不同，23个没有参考文献分配。MethylFLYA比其他算法分配更多的甲基，平均错误率为1%，运行时间为0.4-1.2小时，可以处理任意同位素标记模式和其他类型的NMR光谱数据。
Isotopically labeled methyl groups provide NMR probes in large, otherwise deuterated proteins. However, the resonance assignment constitutes a bottleneck for broader applicability of methyl-based NMR. Here, we present the automated MethylFLYA method for the assignment of methyl groups that is based on methyl-methyl nuclear Overhauser effect spectroscopy (NOESY) peak lists. MethylFLYA is applied to five proteins (28-358 kDa) comprising a total of 708 isotope-labeled methyl groups, of which 612 contribute NOESY cross peaks. MethylFLYA confidently assigns 488 methyl groups, i.e. 80% of those with NOESY data. Of these, 459 agree with the reference, 6 were different, and 23 were without reference assignment. MethylFLYA assigns significantly more methyl groups than alternative algorithms, has an average error rate of 1%, modest runtimes of 0.4-1.2 h, and can handle arbitrary isotope labeling patterns and data from other types of NMR spectra.