Identification of RNA Base Pairs and Complete Assignment of Nucleobase Resonances by Proton-Detected Solid-State NMR Spectroscopy at 100 kHz MAS

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Autoren

  • Philipp Innig Aguion
  • John Kirkpatrick
  • Teresa Carlomagno
  • Alexander Marchanka

Externe Organisationen

  • Helmholtz-Zentrum für Infektionsforschung GmbH (HZI)
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Details

Titel in ÜbersetzungIdentifizierung von RNA-Basenpaaren und vollständige Zuordnung von Nukleobasen-Resonanzen durch Protonen-detektierte Festkörper-NMR-Spektroskopie bei MAS Geschwindigkeiten von 100 kHz
OriginalspracheEnglisch
Seiten (von - bis)23903-23910
Seitenumfang8
FachzeitschriftAngewandte Chemie - International Edition
Jahrgang60
Ausgabenummer44
Frühes Online-Datum28 Sept. 2021
PublikationsstatusVeröffentlicht - 25 Okt. 2021

Abstract

Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.

ASJC Scopus Sachgebiete

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Identification of RNA Base Pairs and Complete Assignment of Nucleobase Resonances by Proton-Detected Solid-State NMR Spectroscopy at 100 kHz MAS. / Aguion, Philipp Innig; Kirkpatrick, John; Carlomagno, Teresa et al.
in: Angewandte Chemie - International Edition, Jahrgang 60, Nr. 44, 25.10.2021, S. 23903-23910.

Publikation: Beitrag in FachzeitschriftArtikelForschungPeer-Review

Aguion PI, Kirkpatrick J, Carlomagno T, Marchanka A. Identification of RNA Base Pairs and Complete Assignment of Nucleobase Resonances by Proton-Detected Solid-State NMR Spectroscopy at 100 kHz MAS. Angewandte Chemie - International Edition. 2021 Okt 25;60(44):23903-23910. Epub 2021 Sep 28. doi: https://doi.org/10.1002/anie.202107263, 10.1002/ange.202107263, 10.15488/12454
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abstract = "Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.",
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AU - Aguion, Philipp Innig

AU - Kirkpatrick, John

AU - Carlomagno, Teresa

AU - Marchanka, Alexander

N1 - Funding Information: This work was supported by the Deutsche Forschungsgemeinschaft (DFG grant CA294/21‐1 to TC). Open Access funding enabled and organized by Projekt DEAL.

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N2 - Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.

AB - Knowledge of RNA structure, either in isolation or in complex, is fundamental to understand the mechanism of cellular processes. Solid-state NMR (ssNMR) is applicable to high molecular-weight complexes and does not require crystallization; thus, it is well-suited to study RNA as part of large multicomponent assemblies. Recently, we solved the first structures of both RNA and an RNA-protein complex by ssNMR using conventional 13C- and 15N-detection. This approach is limited by the severe overlap of the RNA peaks together with the low sensitivity of multidimensional experiments. Here, we overcome the limitations in sensitivity and resolution by using 1H-detection at fast MAS rates. We develop experiments that allow the identification of complete nucleobase spin-systems together with their site-specific base pair pattern using sub-milligram quantities of one uniformly labelled RNA sample. These experiments provide rapid access to RNA secondary structure by ssNMR in protein-RNA complexes of any size.

KW - H detection

KW - base-pair pattern

KW - RNA structure

KW - RNA-protein complex

KW - solid-state NMR spectroscopy

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