Studying Intrinsically Disordered Proteins under True In Vivo Conditions by Combined Cross‐Polarization and Carbonyl‐Detection NMR Spectroscopy |
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| Authors: | Dr. Juan Lopez Dr. Robert Schneider Dr. Francois‐Xavier Cantrelle Dr. Isabelle Huvent Dr. Guy Lippens |
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| Affiliation: | 1. Université de Lille, CNRS, UMR 8576, UGSF—Unité de Glycobiologie Structurale et Fonctionnelle, Lille, France;2. Departamento de Ciencias-Quimica, Pontificia Universidad Catolica del Peru, Lima, Peru;3. Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, CNRS, INRA, INSA Toulouse, Toulouse, France |
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| 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. 13C 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. |
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| Keywords: | α -synuclein carbon detection cross-polarization in-cell NMR spectroscopy intrinsically disordered proteins |
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