Dynamic nuclear polarization on a hybridized hammerhead ribozyme: An explorative study of RNA folding and direct DNP with a paramagnetic metal ion cofactor.
Journal
Solid state nuclear magnetic resonance
ISSN: 1527-3326
Titre abrégé: Solid State Nucl Magn Reson
Pays: Netherlands
ID NLM: 9306181
Informations de publication
Date de publication:
09 2019
09 2019
Historique:
received:
15
03
2019
revised:
16
04
2019
accepted:
16
04
2019
pubmed:
12
5
2019
medline:
26
6
2020
entrez:
12
5
2019
Statut:
ppublish
Résumé
While uniform isotope labeling of ribonucleic acids (RNA) can simply and efficiently be achieved by in-vitro transcription, the specific introduction of nucleotides in larger constructs is non-trivial and often ineffective. Here, we demonstrate how a medium-sized (67-mer), biocatalytically relevant RNA (hammerhead ribozyme, HHRz) can be formed by spontaneous hybridization of two differently isotope-labeled strands, each individually synthesized by in-vitro transcription. This allows on the one hand for a significant reduction in the number of isotope-labeled nucleotides and thus spectral overlap particularly under magic-angle spinning (MAS) dynamic nuclear polarization (DNP) NMR conditions, on the other hand for orthogonal
Identifiants
pubmed: 31078101
pii: S0926-2040(19)30045-1
doi: 10.1016/j.ssnmr.2019.04.005
pii:
doi:
Substances chimiques
Coenzymes
0
RNA, Catalytic
0
hammerhead ribozyme
0
Manganese
42Z2K6ZL8P
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
21-30Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.