Optimal design of adaptively sampled NMR experiments for measurement of methyl group dynamics with application to a ribosome-nascent chain complex
Optimal design of adaptively sampled NMR experiments for measurement of methyl group dynamics with application to a ribosome-nascent chain complex
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DOI：
10.1101/2020.10.12.336511
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发表时间：
2020-10
期刊：
影响因子：
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通讯作者：
C. Waudby;Charles Burridge;J. Christodoulou
中科院分区：
文献类型：
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作者：
C. Waudby;Charles Burridge;J. Christodoulou
交叉相关的核自旋弛豫的NMR测量提供了强大的探针的多肽动力学和旋转扩散，免费的贡献，由于化学交换或与外部自旋的相互作用。在这里，我们报告了一个灵敏度优化的脉冲序列的发展，用于测量甲基自旋系统中的交叉相关弛豫的差分弛豫内的13 C多重峰的跃迁分析。我们描述了应用最优设计理论来实现一个实时的“在飞”自适应采样方案，最大限度地提高测量的速率常数的准确性。使用这种方法获得的灵敏度的增加，使，第一次，定量测量的旋转扩散内的折叠状态的FLN 5细丝结构域的捕获核糖体新生链复合物，并可用于放置强限制域和核糖体表面之间的相互作用。
NMR measurements of cross-correlated nuclear spin relaxation provide powerful probes of polypeptide dynamics and rotational diffusion, free from contributions due to chemical exchange or interactions with external spins. Here, we report on the development of a sensitivity-optimized pulse sequence for the measurement of cross-correlated relaxation in methyl spin systems by analysis of the differential relaxation of transitions within the 13C multiplet. We describe the application of optimal design theory to implement a real-time ‘on-the-fly’ adaptive sampling scheme that maximizes the accuracy of the measured rate constants. The increase in sensitivity obtained using this approach enables, for the first time, quantitative measurements of rotational diffusion within folded states of translationally-arrested ribosome–nascent chain complexes of the FLN5 filamin domain, and can be used to place strong limits on interactions between the domain and the ribosome surface.