title: Studying Intrinsically Disordered Proteins under True In Vivo Conditions by Combined Cross-Polarization and Carbonyl-Detection NMR Spectroscopy.
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.
authors.affiliation: Université de Lille, CNRS, UMR 8576, UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France. juan.lopez@pucp.edu.pe.|Departamento de Ciencias-Quimica, Pontificia Universidad Catolica del Peru, Av. Universitaria 1801, Lima, 32, Peru. juan.lopez@pucp.edu.pe.
authors.forename: Juan
authors.identifier: 0000-0002-5390-6610
authors.initials: J
authors.lastname: Lopez
authors.affiliation: Université de Lille, CNRS, UMR 8576, UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France.
authors.forename: Robert
authors.identifier: 0000-0003-3279-5365
authors.initials: R
authors.lastname: Schneider
authors.affiliation: Université de Lille, CNRS, UMR 8576, UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France.
authors.forename: Francois-Xavier
authors.initials: FX
authors.lastname: Cantrelle
authors.affiliation: Université de Lille, CNRS, UMR 8576, UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Huvent
authors.affiliation: Université de Lille, CNRS, UMR 8576, UGSF-Unité de Glycobiologie Structurale et Fonctionnelle, 59000, Lille, France. glippens@insa-toulouse.fr.|Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, Université de Toulouse, CNRS, INRA, INSA Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France. glippens@insa-toulouse.fr.
authors.forename: Guy
authors.identifier: 0000-0002-8236-0901
authors.initials: G
authors.lastname: Lippens
mesh_terms: D001419:Bacteria; D064267:Intrinsically Disordered Proteins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009584:Nitrogen / Q000737:chemistry; D050050:Protein Carbonylation; D051844:alpha-Synuclein / Q000737:chemistry*
chemical_list: D064267:Intrinsically Disordered Proteins; D051844:alpha-Synuclein; D009584:Nitrogen
keywords: carbon detection; cross-polarization; in-cell NMR spectroscopy; intrinsically disordered proteins; α-synuclein