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Complex formation of the tetracycline-binding aptamer investigated by specific cross-relaxation under DNP.

Aladin, Victoria ; Vogel, Marc ; Binder, Robert ; Burghardt, Irene ; Suess, Beatrix ; Corzilius, Björn (2019):
Complex formation of the tetracycline-binding aptamer investigated by specific cross-relaxation under DNP.
In: Angewandte Chemie (International ed. in English), 58 (15), pp. 4863-4868. ISSN 1521-3773,
DOI: 10.1002/anie.201811941,
[Article]

Abstract

While dynamic nuclear polarization (DNP) under magic-angle spinning (MAS) is generally a powerful method capable of greatly enhancing the sensitivity of solid-state NMR, the displacement of nuclear spin polarization far from thermal equilibrium (i.e., hyperpolarization) also gives rise to peculiar spin dynamics. Here, we elucidate how Specific Cross-Relaxation Enhancement by Active Motions under DNP (SCREAM-DNP) can be utilized in order to selectively obtain MAS NMR spectra of an RNA aptamer in a tightly bound complex with a methyl-bearing ligand (tetracycline) due to the effective CH₃-reorientation at an optimized sample temperature of ~160 K. We demonstrate that SCREAM-DNP is able to spectrally isolate the complex from non-bound species within an RNA mixture and perform a competition assay between the aptamer and a mutant with compromised binding affinity. Finally, we discuss variations in molecular structure and methyl dynamics as observed by SCREAM-DNP between free tetracycline and when bound in a complex with RNA.

Item Type: Article
Erschienen: 2019
Creators: Aladin, Victoria ; Vogel, Marc ; Binder, Robert ; Burghardt, Irene ; Suess, Beatrix ; Corzilius, Björn
Title: Complex formation of the tetracycline-binding aptamer investigated by specific cross-relaxation under DNP.
Language: English
Abstract:

While dynamic nuclear polarization (DNP) under magic-angle spinning (MAS) is generally a powerful method capable of greatly enhancing the sensitivity of solid-state NMR, the displacement of nuclear spin polarization far from thermal equilibrium (i.e., hyperpolarization) also gives rise to peculiar spin dynamics. Here, we elucidate how Specific Cross-Relaxation Enhancement by Active Motions under DNP (SCREAM-DNP) can be utilized in order to selectively obtain MAS NMR spectra of an RNA aptamer in a tightly bound complex with a methyl-bearing ligand (tetracycline) due to the effective CH₃-reorientation at an optimized sample temperature of ~160 K. We demonstrate that SCREAM-DNP is able to spectrally isolate the complex from non-bound species within an RNA mixture and perform a competition assay between the aptamer and a mutant with compromised binding affinity. Finally, we discuss variations in molecular structure and methyl dynamics as observed by SCREAM-DNP between free tetracycline and when bound in a complex with RNA.

Journal or Publication Title: Angewandte Chemie (International ed. in English)
Journal volume: 58
Number: 15
Divisions: 10 Department of Biology
10 Department of Biology > Synthetic Genetic Circuits
Date Deposited: 18 Feb 2019 12:56
DOI: 10.1002/anie.201811941
Identification Number: pmid:30768874
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