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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 international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201308927
Subject(s) - magic angle spinning , deuterium , nuclear magnetic resonance spectroscopy , chemistry , spectroscopy , excitation , excited state , resonance (particle physics) , nuclear magnetic resonance , analytical chemistry (journal) , solid state nuclear magnetic resonance , atomic physics , stereochemistry , physics , chromatography , quantum mechanics
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.