Spectral editing of alanine, serine, and threonine in uniformly labeled proteins based on frequency-selective homonuclear recoupling in solid-state NMR
Spectral editing of alanine, serine, and threonine in uniformly labeled proteins based on frequency-selective homonuclear recoupling in solid-state NMR
复制标题
基于固态 NMR 中的频率选择性同核重偶联，对统一标记的蛋白质中的丙氨酸、丝氨酸和苏氨酸进行光谱编辑
DOI：
10.1007/s10858-021-00367-9
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发表时间：
2021-04-22
影响因子：
2.7
通讯作者：
Yang，Jun
中科院分区：
文献类型：
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作者：
Xiao，Hang;Zhang，Zhengfeng;Yang，Jun
光谱编辑对于简化蛋白质的拥挤固态NMR光谱至关重要。基于我们最近的频率选择性同源重偶联序列，引入新技术来编辑中等魔角旋转（MAS）下均匀标记的蛋白质的13 C-13 C相关性[Zhang et al.，《物理化学杂志》2020，11，8077-8083]。通过选择性13 C α-13 C β双量子滤波（DQF），选择性地滤除丙氨酸、丝氨酸或苏氨酸残基的信号。丙氨酸残基的13 C α-13 C β相关性的选择性建立，效率高达偶极辅助旋转共振（DARR）的约1.8倍。该技术在2D/3D NCCX实验中显示，并应用于均匀13 C，15 N标记的水通道蛋白Z（AqpZ）膜蛋白，证明了它们在生物固态NMR中简化光谱分析的潜力。
Spectral editing is crucial to simplify the crowded solid-state NMR spectra of proteins. New techniques are introduced to edit13C-13C correlations of uniformly labeled proteins under moderate magic-angle spinning (MAS), based on our recent frequency-selective homonuclear recoupling sequences [Zhang et al., J. Phys. Chem. Lett. 2020, 11, 8077–8083]. The signals of alanine, serine, or threonine residues are selected out by selective13Cα-13Cβ double-quantum filtering (DQF). The13Cα-13Cβ correlations of alanine residues are selectively established with efficiency up to ~ 1.8 times that by dipolar-assisted rotational resonance (DARR). The techniques are shown in 2D/3D NCCX experiments and applied to the uniformly13C,15N labeled Aquaporin Z (AqpZ) membrane protein, demonstrating their potential to simplify spectral analyses in biological solid-state NMR.