19F Paramagnetic Relaxation-Based NMR for Quaternary Structural Restraints of Ion Channels.,ACS Chemical Biology

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19F Paramagnetic Relaxation-Based NMR for Quaternary Structural Restraints of Ion Channels.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2019-09-18 , DOI: 10.1021/acschembio.9b00692
Vasyl Bondarenko ₁ , Marta M Wells ₁ , Qiang Chen ₁ , Kevin C Singewald ₂ , Sunil Saxena ₂ , Yan Xu _{1,

3,

4,

5} , Pei Tang _{1,

3,

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Affiliation

Quaternary distance restraints are essential to define the three-dimensional structures of protein assemblies. These distances often fall within a range of 10-18 Å, which challenges the high and low measurement limits of conventional nuclear magnetic resonance (NMR) and double electron-electron resonance electron spin resonance spectroscopies. Here, we report the use of 19F paramagnetic relaxation enhancement (PRE) NMR in combination with 19F/paramagnetic labeling to equivalent sites in different subunits of a protein complex in micelles to determine intersubunit distances. The feasibility of this strategy was evaluated on a pentameric ligand-gated ion channel, for which we found excellent agreement of the 19F PRE NMR results with previous structural information. The study suggests that 19F PRE NMR is a viable tool in extracting distance restraints to define quaternary structures.

中文翻译：

基于19F顺磁性弛豫的NMR用于离子通道的四级结构约束。

四级距离限制对于定义蛋白质装配体的三维结构至关重要。这些距离通常在10-18Å的范围内，这挑战了常规核磁共振（NMR）和双电子-电子共振电子自旋共振光谱仪的高和低测量极限。在这里，我们报告19F顺磁性弛豫增强（PRE）NMR与19F /顺磁性标记结合使用，以胶束中蛋白质复合物的不同亚基的等效位点确定亚基间距离。在五聚体配体门控离子通道上评估了该策略的可行性，为此，我们发现19F PRE NMR结果与先前的结构信息极为吻合。

更新日期：2019-09-18

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