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Evaluation of 15N-detected H–N correlation experiments on increasingly large RNAs
Ist Teil von
Journal of biomolecular NMR, 2017-09, Vol.69 (1), p.31-44
Ort / Verlag
Dordrecht: Springer Netherlands
Erscheinungsjahr
2017
Link zum Volltext
Quelle
SpringerLink
Beschreibungen/Notizen
Recently,
15
N-detected multidimensional NMR experiments have been introduced for the investigation of proteins. Utilization of the slow transverse relaxation of nitrogen nuclei in a
15
N-TROSY experiment allowed recording of high quality spectra for high molecular weight proteins, even in the absence of deuteration. Here, we demonstrate the applicability of three
15
N-detected H–N correlation experiments (TROSY, BEST-TROSY and HSQC) to RNA. With the newly established
15
N-detected BEST-TROSY experiment, which proves to be the most sensitive
15
N-detected H–N correlation experiment, spectra for five RNA molecules ranging in size from 5 to 100 kDa were recorded. These spectra yielded high resolution in the
15
N-dimension even for larger RNAs since the increase in line width with molecular weight is more pronounced in the
1
H- than in the
15
N-dimension. Further, we could experimentally validate the difference in relaxation behavior of imino groups in AU and GC base pairs. Additionally, we showed that
15
N-detected experiments theoretically should benefit from sensitivity and resolution advantages at higher static fields but that the latter is obscured by exchange dynamics within the RNAs.