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. 2020 Oct 21;11:5317. doi: 10.1038/s41467-020-19108-x

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Conventional (100 ms mixing) vs b HMT NOESY spectra (17 loops with a 20 Hz inversion pulse followed by 40 ms mixing each) recorded on the 14mer on top. Both 2D spectra show the imino resonances placed along the vertical direction; for maximizing its efficiency, however, the conventional spectrum was acquired with the imino resonances along the direct domain, and was flipped in this graph to match the HMT data (otherwise, if encoding the iminos in t₁, the conventional experiment showed no peaks altogether). As this was an ¹⁵N-labeled sample, looped inversions/mixing periods were preferred over continuous saturations: this avoided a constant ¹⁵N decoupling during the saturation period, which would have led to sample heating. Assignments are as labeled in the spectra; cross peaks labeled with asterisk are not visible at the given contour level. To better appreciate the enhancements, c and d show various 1D slices extracted from a and b, respectively. Shown in blue for the HMT NOESY are traces addressing U7 and U8 with a stronger (40 Hz continuous) saturation HMT scheme, revealing new correlations despite these sites’ >30 s⁻¹ solvent exchange rates. All spectra were acquired at 1 GHz using a Bruker Avance Neo spectrometer equipped with a TCI cryoprobe.