High-Yield Spin Labeling of Long RNAs for Electron Paramagnetic Resonance Spectroscopy

Mark Kerzhner; Hideto Matsuoka; Christine Wuebben; Michael Famulok; Olav Schiemann

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High-Yield Spin Labeling of Long RNAs for Electron Paramagnetic Resonance Spectroscopy

机译：用于电子顺磁共振光谱的长RNA的高产旋转标记

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Site-directed spin labeling is a powerful tool for investigating the conformation and dynamics of biomacromolecules such as RNA. Here we introduce a spin labeling strategy based on click chemistry in solution that, in combination with enzymatic ligation, allows highly efficient labeling of complex and long RNAs with short reaction times and suppressed RNA degradation. With this approach, a 34-nucleotide aptamer domain of the preQ1 riboswitch and an 81-nucleotide TPP riboswitch aptamer could be labeled with two labels in several positions. We then show that conformations of the preQ1 aptamer and its dynamics can be monitored in the absence and presence of Mg~(2+) and a preQ1 ligand by continuous wave electron paramagnetic resonance spectroscopy at room temperature and pulsed electron–electron double resonance spectroscopy (PELDOR or DEER) in the frozen state.

机译：站点定向的旋转标签是一种强大的工具，用于研究生物致摩托（如RNA）的构象和动态。在这里，我们介绍基于单击化学的旋转标签策略，在溶液中，与酶联结合，允许具有短反应时间和抑制RNA降解的复杂和长RNA的高效标记。利用这种方法，可以用几个位置用两个标签标记PRIQ1核糖开关和81核苷酸TPP核糖织物容适体的34核苷酸适体结构域。然后，我们可以在室温下通过连续波电子顺磁共振谱通过在室温和脉冲电子 - 电子双共振光谱（在冻结状态下的乖巧或鹿。

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来源
《Biochemistry》 |2018年第20期|共9页
作者
Mark Kerzhner; Hideto Matsuoka; Christine Wuebben; Michael Famulok; Olav Schiemann;
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作者单位

Life &

Medical Sciences Institute Chemical Biology &

Medicinal Chemistry Unit c/o Kekulé-Institut für Organische Chemie und Biochemie University of Bonn;

Institute of Physical and Theoretical Chemistry University of Bonn;

Institute of Physical and Theoretical Chemistry University of Bonn;

Life &

Medical Sciences Institute Chemical Biology &

Medicinal Chemistry Unit c/o Kekulé-Institut für Organische Chemie und Biochemie University of Bonn;

Institute of Physical and Theoretical Chemistry University of Bonn;

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正文语种 eng
中图分类生物化学;
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专利

1. High-Yield Spin Labeling of Long RNAs for Electron Paramagnetic Resonance Spectroscopy [J] . Mark Kerzhner, Hideto Matsuoka, Christine Wuebben, Biochemistry . 2018,第20期

机译：用于电子顺磁共振光谱的长RNA的高产旋转标记
2. Site‐directed spin label electron paramagnetic resonance spectroscopy as a probe of conformational dynamics in the Fe(III) “ locked‐off locked‐off ” state of the CO‐sensing transcription factor CooA [J] . Hines Judy P., Dent Matthew R., Stevens Daniel J., Protein Science: A Publication of the Protein Society . 2018,第9期

机译：站点定向的旋转标签电子顺磁共振谱作为共同传感转录因子COOA的FE（III）“锁定锁定”状态的组合动力学探针
3. Determination of the Complete Spin Density Distribution in C-13-Labeled Protein-Bound Radical Intermediates Using Advanced 2D Electron Paramagnetic Resonance Spectroscopy and Density Functional Theory [J] . Taguchi Alexander T., OMalley Patrick J., Wraight Colin A., The journal of physical chemistry, B. Condensed matter, materials, surfaces, interfaces & biophysical . 2017,第44期

机译：使用先进的2D电子顺笔谐振谱和密度泛函理论测定C-13标记蛋白质结合的基团中间体的完全自旋密度分布
4. We have investigated the interface and oxide defects in SiC/SiO_2 by electron paramagnetic resonance spectroscopy (EPR) using oxidised porous SiC of 4H and 6H polytypes. Two paramagnetic defects generated by 1000 deg C furnace oxidation in dry oxygen [C] . Krishna C. Saraswat, Chi On Chui, Paul C. Mclntyre, the International Symposium of Electrochemical Society . 2003

机译：通过4H和6H多型，通过电子顺磁共振光谱（EPR）研究了SiC / SiO_2中的界面和氧化物缺陷。在干氧中1000°C炉氧化产生的两个顺磁缺陷
5. Electron paramagnetic resonance spectroscopy of spin-labeled RNA: An emerging tool for the elucidation of RNA structure and dynamics. [D] . Edwards, Thomas Eugene. 2003

机译：自旋标记RNA的电子顺磁共振波谱：阐明RNA结构和动力学的新兴工具。
6. A novel 5 displacement spin-labeling technique for electron paramagnetic resonance spectroscopy of RNA. [O] . J C Macosko, M S Pio, I Tinoco Jr, 1999

机译：RNA的电子顺磁共振波谱的一种新型5位移自旋标记技术。
7. Studying RNA Using Site-Directed Spin-Labeling and Continuous-Wave Electron Paramagnetic Resonance Spectroscopy [O] . Xiaojun Zhang, Pavol Cekan, Snorri Th. Sigurdsson, 2009

机译：使用位点定向的旋转标记和连续波电子顺磁共振谱学研究RNA

High-Yield Spin Labeling of Long RNAs for Electron Paramagnetic Resonance Spectroscopy

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