Evaluation of 15N-detected H-N correlation experiments on increasingly large RNAs.
J Biomol NMR
; 69(1): 31-44, 2017 Sep.
Article
em En
| MEDLINE
| ID: mdl-28879611
ABSTRACT
Recently, 15N-detected multidimensional NMR experiments have been introduced for the investigation of proteins. Utilization of the slow transverse relaxation of nitrogen nuclei in a 15N-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 15N-detected H-N correlation experiments (TROSY, BEST-TROSY and HSQC) to RNA. With the newly established 15N-detected BEST-TROSY experiment, which proves to be the most sensitive 15N-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 15N-dimension even for larger RNAs since the increase in line width with molecular weight is more pronounced in the 1H- than in the 15N-dimension. Further, we could experimentally validate the difference in relaxation behavior of imino groups in AU and GC base pairs. Additionally, we showed that 15N-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.
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MEDLINE
Assunto principal:
RNA
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Ressonância Magnética Nuclear Biomolecular
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Isótopos de Nitrogênio
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article