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J Biomol NMR. 2018 Mar;70(3):177-185. doi: 10.1007/s10858-018-0168-3. Epub 2018 Mar 3.

Detection of side-chain proton resonances of fully protonated biosolids in nano-litre volumes by magic angle spinning solid-state NMR.

Author information

1
Institute of Chemistry & Biology of Membranes & Nanoobjects, (UMR5248 CBMN), CNRS, Université Bordeaux, Institut Européen de Chimie et Biologie, 33600, Pessac, France.
2
JEOL RESONANCE Inc., Musashino, Akishima, Tokyo, 196-8558, Japan.
3
RIKEN CLST-JEOL Collaboration Center, Tsurumi, Yokohama, Kanagawa, 230-0045, Japan.
4
Department of Chemistry, Indian Institute of Technology Ropar, Rupnagar, India.
5
Institut de Biochimie et de Génétique Cellulaire, (UMR 5095 IBGC), CNRS, Université Bordeaux, 33077, Bordeaux, France.
6
JEOL RESONANCE Inc., Musashino, Akishima, Tokyo, 196-8558, Japan. yunishiy@jeol.co.jp.
7
RIKEN CLST-JEOL Collaboration Center, Tsurumi, Yokohama, Kanagawa, 230-0045, Japan. yunishiy@jeol.co.jp.
8
Institute of Chemistry & Biology of Membranes & Nanoobjects, (UMR5248 CBMN), CNRS, Université Bordeaux, Institut Européen de Chimie et Biologie, 33600, Pessac, France. b.habenstein@iecb.u-bordeaux.fr.
9
Institute of Chemistry & Biology of Membranes & Nanoobjects, (UMR5248 CBMN), CNRS, Université Bordeaux, Institut Européen de Chimie et Biologie, 33600, Pessac, France. a.loquet@iecb.u-bordeaux.fr.

Abstract

We present a new solid-state NMR proton-detected three-dimensional experiment dedicated to the observation of protein proton side chain resonances in nano-liter volumes. The experiment takes advantage of very fast magic angle spinning and double quantum 13C-13C transfer to establish efficient (H)CCH correlations detected on side chain protons. Our approach is demonstrated on the HET-s prion domain in its functional amyloid fibrillar form, fully protonated, with a sample amount of less than 500 µg using a MAS frequency of 70 kHz. The majority of aliphatic and aromatic side chain protons (70%) are observable, in addition to Hα resonances, in a single experiment providing a complementary approach to the established proton-detected amide-based multidimensional solid-state NMR experiments for the study and resonance assignment of biosolid samples, in particular for aromatic side chain resonances.

KEYWORDS:

Amyloid fibrils; Protein NMR; Proton detection; Solid-state NMR; Very fast MAS

PMID:
29502224
DOI:
10.1007/s10858-018-0168-3
[Indexed for MEDLINE]

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