Sparse C-13 labelling for solid-state NMR studies of P. pastoris expressed eukaryotic seven-transmembrane proteins
Sparse C-13 labelling for solid-state NMR studies of P. pastoris expressed eukaryotic seven-transmembrane proteins
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用于巴斯德毕赤酵母表达的真核七跨膜蛋白的固态 NMR 研究的稀疏 C-13 标记
DOI：
10.1007/s10858-016-0033-1
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发表时间：
2016
影响因子：
2.7
通讯作者：
Wang Shenlin
中科院分区：
文献类型：
--
作者：
Liu Jing;Liu Chang;Fan Ying;Munro Rachel A.;Ladizhansky Vladimir;Brown Leonid S.;Wang Shenlin
我们展示了一种新的用于甲基营养酵母的sparse13C标记方法。巴斯德酵母表达系统，迈向真核生物膜蛋白的固态核磁共振研究。标记方案是通过在表达培养基中共同利用天然丰度的甲醇和13c标记的甘油作为碳源来实现的。该策略将光谱分辨率提高了1.5倍，显示了位点特异性标记模式，并且具有收集远距离距离限制的优势，可用于固体核磁共振测定大型真核膜蛋白的结构。
We demonstrate a novel sparse13C labelling approach for methylotrophic yeastP. pastorisexpression system, towards solid-state NMR studies of eukaryotic membrane proteins. The labelling scheme was achieved by co-utilizing natural abundance methanol and specifically13C labelled glycerol as carbon sources in the expression medium. This strategy improves the spectral resolution by 1.5 fold, displays site-specific labelling patterns, and has advantages for collecting long-range distance restraints for structure determination of large eukaryotic membrane proteins by solid-state NMR.