Schnieders, R, Richter, C, Warhaut, S et al. (10 more authors) (2017) Evaluation of (15)N-detected H-N correlation experiments on increasingly large RNAs. Journal of Biomolecular NMR, 69 (1). pp. 31-44. ISSN 1573-5001
Abstract
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.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2017, Springer. This is an author produced version of a paper published in Journal of Biomolecular NMR. Uploaded in accordance with the publisher's self-archiving policy. The final publication is available at http://dx.doi.org/10.1007/s10858-017-0132-7 |
Keywords: | 15N direct detection; BEST-TROSY; Exchange; Field dependence; Line width analysis; RNA |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > NMR (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 19 Oct 2017 11:45 |
Last Modified: | 06 Sep 2018 00:39 |
Status: | Published |
Publisher: | Springer Nature |
Identification Number: | 10.1007/s10858-017-0132-7 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:122778 |