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)
doi: 10.1002/anie.201811941
Artikel, Bibliographie
Kurzbeschreibung (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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2019 |
| Autor(en): | Aladin, Victoria ; Vogel, Marc ; Binder, Robert ; Burghardt, Irene ; Suess, Beatrix ; Corzilius, Björn |
| Art des Eintrags: | Bibliographie |
| Titel: | Complex formation of the tetracycline-binding aptamer investigated by specific cross-relaxation under DNP. |
| Sprache: | Englisch |
| Publikationsjahr: | 1 April 2019 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Angewandte Chemie (International ed. in English) |
| Jahrgang/Volume einer Zeitschrift: | 58 |
| (Heft-)Nummer: | 15 |
| DOI: | 10.1002/anie.201811941 |
| Kurzbeschreibung (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. |
| ID-Nummer: | pmid:30768874 |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") |
| Hinterlegungsdatum: | 18 Feb 2019 12:56 |
| Letzte Änderung: | 29 Apr 2019 06:54 |
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