Versatile Trityl Spin Labels for Nanometer Distance Measurements on Biomolecules In Vitro and within Cells
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- 作者：Dipl.-Chem. J. Jacques Jassoy ; Dipl.-Chem. Andreas Berndhä ; user ; B. Sc. Fraser Duthie ; B. Sc. Sebastian P. Kü ; hn ; Dr. Gregor Hagelueken and Prof. Dr. Olav Schiemann
- 刊名：Angewandte Chemie International Edition
- 出版年：2017
- 出版时间：January 2, 2017
- 年：2017
- 卷：56
- 期：1
- 页码：177-181
- 全文大小：1028K
- ISSN：1521-3773
文摘
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.