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De Novo 3D Structure Determination from Sub‐milligram Protein Samples by Solid‐State 100 kHz MAS NMR Spectroscopy
Author(s) -
Agarwal Vipin,
Penzel Susanne,
Szekely Kathrin,
Cadalbert Riccardo,
Testori Emilie,
Oss Andres,
Past Jaan,
Samoson Ago,
Ernst Matthias,
Böckmann Anja,
Meier Beat H.
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.201405730
Subject(s) - spectroscopy , solid state nuclear magnetic resonance , nuclear magnetic resonance spectroscopy , chemistry , resolution (logic) , magic angle spinning , proton , crystallography , protein structure , analytical chemistry (journal) , membrane protein , magic angle , membrane , nuclear magnetic resonance , chromatography , physics , stereochemistry , biochemistry , quantum mechanics , artificial intelligence , computer science
Solid‐state NMR spectroscopy is an emerging tool for structural studies of crystalline, membrane‐associated, sedimented, and fibrillar proteins. A major limitation for many studies is still the large amount of sample needed for the experiments, typically several isotopically labeled samples of 10–20 mg each. Here we show that a new NMR probe, pushing magic‐angle sample rotation to frequencies around 100 kHz, makes it possible to narrow the proton resonance lines sufficiently to provide the necessary sensitivity and spectral resolution for efficient and sensitive proton detection. Using restraints from such spectra, a well‐defined de novo structure of the model protein ubiquitin was obtained from two samples of roughly 500 μg protein each. This proof of principle opens new avenues for structural studies of proteins available in microgram, or tens of nanomoles, quantities that are, for example, typically achieved for eukaryotic membrane proteins by in‐cell or cell‐free expression.