PHRONESIS: A One-Shot Approach for Sequential Assignment of Protein Resonances by Ultrafast MAS Solid-State NMR Spectroscopy.
PHRONESIS: A One-Shot Approach for Sequential Assignment of Protein Resonances by Ultrafast MAS Solid-State NMR Spectroscopy.
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DOI：
10.1002/cphc.202200127
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发表时间：
2022-07-05
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固态NMR（ssNMR）光谱已成为分析纤维状、膜结合和结晶蛋白质的结构动力学的首选方法，这些蛋白质与其他结构技术无关。近年来，快速魔角旋转下的1H检测和多重获取ssNMR技术推动了复杂生物大分子的结构分析。然而，数据采集和特定谐振分配仍然是该技术的瓶颈。在这里，我们提出了一个全面的多采集实验（PHRONESIS），同时生成多达10个3D 1H检测的ssNMR光谱。PHRONESIS利用宽带传输和选择性脉冲来驱动多个独立的偏振路径。通过结合进化算法和人工智能设计的高保真选择性脉冲实现了特定共振的高选择性激发和去激发。我们用微晶U-13 C，15 N GB 1蛋白证明了这种方法的功效，使用10个3D实验的一个数据集达到100%的共振归属。这项工作中概述的策略为实施新型1H检测多采集ssNMR实验开辟了新的途径，以加速和扩展应用到更大的生物分子系统。 一个全面的固态NMR实验的蛋白质共振顺序分配- PHRONESIS -。
Solid‐state NMR (ssNMR) spectroscopy has emerged as the method of choice to analyze the structural dynamics of fibrillar, membrane‐bound, and crystalline proteins that are recalcitrant to other structural techniques. Recently, 1H detection under fast magic angle spinning and multiple acquisition ssNMR techniques have propelled the structural analysis of complex biomacromolecules. However, data acquisition and resonance‐specific assignments remain a bottleneck for this technique. Here, we present a comprehensive multi‐acquisition experiment (PHRONESIS) that simultaneously generates up to ten 3D 1H‐detected ssNMR spectra. PHRONESIS utilizes broadband transfer and selective pulses to drive multiple independent polarization pathways. High selectivity excitation and de‐excitation of specific resonances were achieved by high‐fidelity selective pulses that were designed using a combination of an evolutionary algorithm and artificial intelligence. We demonstrated the power of this approach with microcrystalline U‐13C,15N GB1 protein, reaching 100 % of the resonance assignments using one data set of ten 3D experiments. The strategy outlined in this work opens up new avenues for implementing novel 1H‐detected multi‐acquisition ssNMR experiments to speed up and expand the application to larger biomolecular systems. A comprehensive solid‐state NMR experiment for sequential assignment of protein resonances – PHRONESIS – is presented.