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Quadruple‐Resonance Magic‐Angle Spinning NMR Spectroscopy of Deuterated Solid Proteins
Author(s) -
Akbey Ümit,
Nieuwkoop Andrew J.,
Wegner Sebastian,
Voreck Anja,
Kunert Britta,
Bandara Priyanga,
Engelke Frank,
Nielsen Niels Chr.,
Oschkinat Hartmut
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201308927
Subject(s) - magic angle spinning , chemistry , nuclear magnetic resonance spectroscopy , deuterium , spectroscopy , excited state , excitation , resonance (particle physics) , solid state nuclear magnetic resonance , nuclear magnetic resonance , analytical chemistry (journal) , crystallography , stereochemistry , atomic physics , physics , chromatography , quantum mechanics
Abstract 1 H‐detected magic‐angle spinning NMR experiments facilitate structural biology of solid proteins, which requires using deuterated proteins. However, often amide protons cannot be back‐exchanged sufficiently, because of a possible lack of solvent exposure. For such systems, using 2 H excitation instead of 1 H excitation can be beneficial because of the larger abundance and shorter longitudinal relaxation time, T 1 , of deuterium. A new structure determination approach, “quadruple‐resonance NMR spectroscopy”, is presented which relies on an efficient 2 H‐excitation and 2 H‐ 13 C cross‐polarization (CP) step, combined with 1 H detection. We show that by using 2 H‐excited experiments better sensitivity is possible on an SH3 sample recrystallized from 30 % H 2 O. For a membrane protein, the ABC transporter ArtMP in native lipid bilayers, different sets of signals can be observed from different initial polarization pathways, which can be evaluated further to extract structural properties.