caa a22 4500 445325186 CHVBK 20180317142718.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9530-4 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9530-4 Extensive de novo solid-state NMR assignments of the 33kDa C-terminal domain of the Ure2 prion [Elektronische Daten] [Birgit Habenstein, Christian Wasmer, Luc Bousset, Yannick Sourigues, Anne Schütz, Antoine Loquet, Beat Meier, Ronald Melki, Anja Böckmann] We present the de novo resonance assignments for the crystalline 33kDa C-terminal domain of the Ure2 prion using an optimized set of five 3D solid-state NMR spectra. We obtained, using a single uniformly 13C, 15N labeled protein sample, sequential chemical-shift information for 74% of the N, Cα, Cβ triples, and for 80% of further side-chain resonances for these spin systems. We describe the procedures and protocols devised, and discuss possibilities and limitations of the assignment of this largest protein assigned today by solid-state NMR, and for which no solution-state NMR shifts were available. A comparison of the NMR chemical shifts with crystallographic data reveals that regions with high crystallographic B-factors are particularly difficult to assign. While the secondary structure elements derived from the chemical shift data correspond mainly to those present in the X-ray crystal structure, we detect an additional helical element and structural variability in the protein crystal, most probably originating from the different molecules in the asymmetric unit, with the observation of doubled resonances in several parts, including entire stretches, of the protein. Our results provide the point of departure towards an atomic-resolution structural analysis of the C-terminal Ure2p domain in the context of the full-length prion fibrils. Springer Science+Business Media B.V., 2011 Prion nationallicence Solid-state NMR nationallicence Fibrils nationallicence Sequential assignment nationallicence Conformational heterogeneity nationallicence Habenstein Birgit Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut Wasmer Christian Harvard Medical School, 02115, Boston, MA, USA aut Bousset Luc Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut Sourigues Yannick Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut Schütz Anne Physical Chemistry, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland aut Loquet Antoine Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut Meier Beat Physical Chemistry, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland aut Melki Ronald Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut Böckmann Anja Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 235-243 0925-2738 51:3<235 2011 51 10858 https://doi.org/10.1007/s10858-011-9530-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9530-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Habenstein Birgit Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut NATIONALLICENCE 700 1- Wasmer Christian Harvard Medical School, 02115, Boston, MA, USA aut NATIONALLICENCE 700 1- Bousset Luc Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut NATIONALLICENCE 700 1- Sourigues Yannick Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut NATIONALLICENCE 700 1- Schütz Anne Physical Chemistry, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Loquet Antoine Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut NATIONALLICENCE 700 1- Meier Beat Physical Chemistry, ETH Zurich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Melki Ronald Laboratoire d'Enzymologie et Biochimie Structurales, UPR 3082 CNRS, Avenue de la Terrasse, 91198, Gif-sur-Yvette, France aut NATIONALLICENCE 700 1- Böckmann Anja Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367, Lyon, France aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 235-243 0925-2738 51:3<235 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer