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. 2019 Nov 4;9:15903. doi: 10.1038/s41598-019-52383-3

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© The Author(s) 2019

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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(A–E) ¹H,¹H contact map for the 1 μs MD relaxed crystal structure of the SH3 domain of α-spectrin for an upper ¹H,¹H distance limit of 5 Å. The maps were calculated for the (A) 5% GlcRAP and (C) Leu/Val labelling scheme, respectively, as well as for an (E) uniformly protonated sample. Full proton back-substitution was also considered for the former in (B) and (D). (F) The probability P to detect at least one ¹H,¹H contact for N adjoining sites assuming a likelihood of p = 0.035 to find a proton at a given site. (G) Methyl region of a ¹H,¹H 2D projection extracted from a ¹H-detected 3D H(H)CH RFDR spectrum (τ_mix = 8 ms) using the 5% GlcRAP labelled SH3 domain¹². The spectrum was recorded at an external magnetic field of 14.1 T (600 MHz) and a MAS frequency of 20 kHz. The H₂O content in the microcrystalline sample was < 10%. Clearly, a number of ¹H,¹H contacts were detected including H^met-H^met, H^met-H^ali, H^met-solvent and H^met-H^N. The figure was generated with Adobe Illustrator CS5 V.15.02.