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Complex formation of the tetracycline-binding aptamer investigated by specific cross-relaxation under DNP.
Angewandte Chemie 2019 Februrary 16
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.
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