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  High-yield spin labeling of long RNAs for electron paramagnetic resonance spectroscopy

Kerzhner, M., Matsuoka, H., Wuebben, C., Famulok, M., & Schiemann, O. (2018). High-yield spin labeling of long RNAs for electron paramagnetic resonance spectroscopy. Biochemistry, 57(20), 2923-2931. doi:10.1021/acs.biochem.8b00040.

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 Creators:
Kerzhner, Mark1, Author
Matsuoka, Hideto1, Author
Wuebben, Christine1, Author
Famulok, Michael2, Author
Schiemann, Olav1, Author
Affiliations:
1External Organizations, ou_persistent22              
2Max Planck Fellow Chemical Biology, Center of Advanced European Studies and Research (caesar), Max Planck Society, Ludwig-Erhard-Allee 2, 53175 Bonn, DE, ou_2173681              

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 Abstract: 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 Mg2+ 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.

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Language(s): eng - English
 Dates: 2018-05-102018-05-22
 Publication Status: Issued
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 Rev. Type: Peer
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Title: Biochemistry
  Abbreviation : Biochemistry
Source Genre: Journal
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Publ. Info: Columbus, Ohio : American Chemical Society
Pages: - Volume / Issue: 57 (20) Sequence Number: - Start / End Page: 2923 - 2931 Identifier: ISSN: 0006-2960
CoNE: https://pure.mpg.de/cone/journals/resource/954925384103