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Studying Intrinsically Disordered Proteins under True In Vivo Conditions by Combined Cross‐Polarization and Carbonyl‐Detection NMR Spectroscopy
Author(s) -
Lopez Juan,
Schneider Robert,
Cantrelle FrancoisXavier,
Huvent Isabelle,
Lippens Guy
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201601850
Subject(s) - chemistry , nuclear magnetic resonance spectroscopy , proton , intrinsically disordered proteins , pulse sequence , spectroscopy , amide , proton nmr , polarization (electrochemistry) , analytical chemistry (journal) , nuclear magnetic resonance , chromatography , organic chemistry , physics , biochemistry , quantum mechanics
Abstract Under physiological conditions, studies of intrinsically disordered proteins (IDPs) by conventional NMR methods based on proton detection are severely limited by fast amide‐proton exchange with water. 13 C detection has been proposed as a solution to the exchange problem, but is hampered by low sensitivity. We propose a new pulse sequence combining proton–nitrogen cross‐polarization and carbonyl detection to record high‐resolution, high‐sensitivity NMR spectra of IDPs under physiological conditions. To demonstrate the efficacy of this approach, we recorded a high‐quality N–CO correlation spectrum of α‐synuclein in bacterial cells at 37 °C.