Publication – Fürtig, Schwalbe, Wöhnert

Evaluation of 15N-detected H-N correlation experiments on increasingly large RNAs.

Schnieders R, Richter C, Warhaut S, de Jesus V, Keyhani S, Duchardt-Ferner E, Keller H, Wöhnert J, Kuhn LT, Breeze AL, Bermel W, Schwalbe H, Fürtig B.
J. Biomol. NMR, 2017 Sep 6.

Full publication here: Evaluation of 15N-detected H–N correlation experiments on increasingly large RNAs

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.