Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells
Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells
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DOI：
10.1002/anie.201609085
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发表时间：
2017-01-02
影响因子：
16.6
通讯作者：
Schiemann, Olav
中科院分区：
文献类型：
--
作者：
Jassoy, J. Jacques;Berndhaeuser, Andreas;Schiemann, Olav
在细胞内条件下测定生物大分子的结构是一项相关但具有挑战性的任务。电子顺磁共振（EPR）距离测量结合定点自旋标记（SDSL）是一个有价值的工具，在这奋进，但通常使用的氮氧化物自旋标记不适合在细胞内测量。相比之下，三芳基甲基（三苯甲基）自由基是高度持久的，表现出长的弛豫时间和窄的光谱宽度。在这里，一个通用的集合的三苯甲基自旋标记和它们的应用程序在体外和细胞内的三苯甲基铁距离测量细胞色素P450蛋白的合成进行了描述。与流行的甲硫基磺酸盐自旋标记（MTSSL）相比，三苯甲基标记显示出类似的标记效率和更好的信噪比（SNR），并且能够成功地进行细胞内测量。
Structure determination of biomacromolecules under in-cell conditions is a relevant yet challenging task. Electron paramagnetic resonance (EPR) distance measurements in combination with site-directed spin labeling (SDSL) are a valuable tool in this endeavor but the usually used nitroxide spin labels are not well-suited for in-cell measurements. In contrast, triarylmethyl (trityl) radicals are highly persistent, exhibit a long relaxation time and a narrow spectral width. Here, the synthesis of a versatile collection of trityl spin labels and their application in in vitro and in-cell trityl-iron distance measurements on a cytochrome P450 protein are described. The trityl labels show similar labeling efficiencies and better signal-to-noise ratios (SNR) as compared to the popular methanethiosulfonate spin label (MTSSL) and enabled a successful in-cell measurement.