A Comparison of Cysteine-Conjugated Nitroxide Spin Labels for Pulse Dipolar EPR Spectroscopy.
A Comparison of Cysteine-Conjugated Nitroxide Spin Labels for Pulse Dipolar EPR Spectroscopy.
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脉冲偶极EPR光谱法的半胱氨酸偶联的氮氧化物自旋标记的比较。
DOI：
10.3390/molecules26247534
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发表时间：
2021-12-13
期刊：
影响因子：
--
通讯作者：
Bode BE
中科院分区：
文献类型：
--
作者：
Ackermann K;Chapman A;Bode BE
结构-功能和材料范式推动了从材料科学到结构生物学对分子和固体的结构和结构异质性的理解。对复杂建筑的功能洞察通常是从一套互补的物理化学方法中获得的。在生物大分子结构的研究中，脉冲偶极电子顺磁共振波谱(PDS)的应用越来越普遍。PDS的主要兴趣是提供长程纳米距离分布，允许识别大分子拓扑结构、验证结构模型和构象转变以及四元络合物的对接。最常见的是，半胱氨酸通过定点突变引入蛋白质结构，并被修饰到自旋标记的侧链上，如稳定的氮氧自由基。在这篇文章中，我们研究了四种不同的商业标记剂通过不同的硫特定反应进行的标记。此外，所获得的距离分布是自旋承载部分之间的距离分布，并且需要通过建模方法与蛋白质结构相关。在这里，我们比较两种不同的方法来模拟所有四个侧链的这些分布。结果表明，最佳标记方法存在显著差异。所有四种自旋标记在标记和纯化的简易性方面都显示出不同。进一步的挑战来自不同的自旋标记侧链的系链长度和旋转体；两者都影响到结构的建模和转换。我们的比较表明，在自旋标记侧基(bis-(2，2，5，5-Tetramethyl-3-imidazoline-1-oxyl-4-yl)二硫化物中具有最短锚链的自旋标记可能被低估，如果相应地优化标记条件，可能会提高PDS结构研究的分辨率。
The structure-function and materials paradigms drive research on the understanding of structures and structural heterogeneity of molecules and solids from materials science to structural biology. Functional insights into complex architectures are often gained from a suite of complementary physicochemical methods. In the context of biomacromolecular structures, the use of pulse dipolar electron paramagnetic resonance spectroscopy (PDS) has become increasingly popular. The main interest in PDS is providing long-range nanometre distance distributions that allow for identifying macromolecular topologies, validating structural models and conformational transitions as well as docking of quaternary complexes. Most commonly, cysteines are introduced into protein structures by site-directed mutagenesis and modified site-specifically to a spin-labelled side-chain such as a stable nitroxide radical. In this contribution, we investigate labelling by four different commercial labelling agents that react through different sulfur-specific reactions. Further, the distance distributions obtained are between spin-bearing moieties and need to be related to the protein structure via modelling approaches. Here, we compare two different approaches to modelling these distributions for all four side-chains. The results indicate that there are significant differences in the optimum labelling procedure. All four spin-labels show differences in the ease of labelling and purification. Further challenges arise from the different tether lengths and rotamers of spin-labelled side-chains; both influence the modelling and translation into structures. Our comparison indicates that the spin-label with the shortest tether in the spin-labelled side-group, (bis-(2,2,5,5-Tetramethyl-3-imidazoline-1-oxyl-4-yl) disulfide, may be underappreciated and could increase the resolution of structural studies by PDS if labelling conditions are optimised accordingly.
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影响因子：
3.4
作者：
Ackermann K;Pliotas C;Valera S;Naismith JH;Bode BE
通讯作者：
Bode BE
旋转对称的同性恋者中的Peldor。
影响因子：
1.7
作者：
Giannoulis A;Ward R;Branigan E;Naismith JH;Bode BE
通讯作者：
Bode BE
影响因子：
1
作者：
Hagelueken, Gregor;Ward, Richard;Naismith, James H.;Schiemann, Olav
通讯作者：
Schiemann, Olav
DOI：
10.1021/acs.jpcb.1c03666
发表时间：
2021-05-27
期刊：
The journal of physical chemistry. B
影响因子：
--
作者：
Ackermann K;Wort JL;Bode BE
通讯作者：
Bode BE
DOI：
10.5194/mr-1-209-2020
发表时间：
2020
期刊：
Magnetic resonance (Gottingen, Germany)
影响因子：
--
作者：
Fábregas Ibáñez L;Jeschke G;Stoll S
通讯作者：
Stoll S