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Angew Chem Int Ed Engl. 2016 Dec 12;55(50):15504-15509. doi: 10.1002/anie.201607084. Epub 2016 Nov 16.

NMR Spectroscopic Assignment of Backbone and Side-Chain Protons in Fully Protonated Proteins: Microcrystals, Sedimented Assemblies, and Amyloid Fibrils.

Author information

1
Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
2
Biomedical Research and Study Centre, Rātsupītes 1, LV1067, Riga, Latvia.
3
Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.
4
School of Chemistry, University of Wollongong, NSW, 2522, Australia.
5
Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
6
Institut de Biochimie et de Génétique Cellulaire (UMR 5095, CNRS -, Université de Bordeaux), 33077, Bordeaux, France.

Abstract

We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100-111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα-Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration.

KEYWORDS:

magic-angle spinning; proton detection; resonance assignment; solid-state NMR spectroscopy

PMID:
27865050
DOI:
10.1002/anie.201607084
[Indexed for MEDLINE]

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