title: NMR Spectroscopic Assignment of Backbone and Side-Chain Protons in Fully Protonated Proteins: Microcrystals, Sedimented Assemblies, and Amyloid Fibrils.
abstract: We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100-111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα-Hα detection block was developed and applied for the sequence-specific backbone and aliphatic side-chain resonance assignment using only 500 μg of sample. These developments accelerate structural studies of biomolecular assemblies available in submilligram quantities without the need of protein deuteration.
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Jan
authors.initials: J
authors.lastname: Stanek
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Loren B
authors.initials: LB
authors.lastname: Andreas
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Kristaps
authors.initials: K
authors.lastname: Jaudzems
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Diane
authors.initials: D
authors.lastname: Cala
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Daniela
authors.initials: D
authors.lastname: Lalli
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bertarello
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Tobias
authors.initials: T
authors.lastname: Schubeis
authors.affiliation: Biomedical Research and Study Centre, Rātsupītes 1, LV1067, Riga, Latvia.
authors.forename: Inara
authors.initials: I
authors.lastname: Akopjana
authors.affiliation: Biomedical Research and Study Centre, Rātsupītes 1, LV1067, Riga, Latvia.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Kotelovica
authors.affiliation: Biomedical Research and Study Centre, Rātsupītes 1, LV1067, Riga, Latvia.
authors.forename: Kaspars
authors.initials: K
authors.lastname: Tars
authors.affiliation: Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pica
authors.affiliation: Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Leone
authors.affiliation: Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126, Naples, Italy.
authors.forename: Delia
authors.initials: D
authors.lastname: Picone
authors.affiliation: School of Chemistry, University of Wollongong, NSW, 2522, Australia.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Xu
authors.affiliation: School of Chemistry, University of Wollongong, NSW, 2522, Australia.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Dixon
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Martinez
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Mélanie
authors.initials: M
authors.lastname: Berbon
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Nadia
authors.initials: N
authors.lastname: El Mammeri
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Abdelmajid
authors.initials: A
authors.lastname: Noubhani
authors.affiliation: Institut de Biochimie et de Génétique Cellulaire (UMR 5095, CNRS -, Université de Bordeaux), 33077, Bordeaux, France.
authors.forename: Sven
authors.initials: S
authors.lastname: Saupe
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Birgit
authors.initials: B
authors.lastname: Habenstein
authors.affiliation: Institute of Chemistry & Biology of Membranes & Nanoobjects (UMR 5248 CBMN - CNRS, University of Bordeaux, Bordeaux INP), All. Geoffroy Saint-Hillaire, 33600, Pessac, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.affiliation: Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (UMR 5280 - CNRS, ENS Lyon, UCB Lyon 1), Université de Lyon, 5 rue de la Doua, 69100, Villeurbanne, France.
authors.forename: Guido
authors.initials: G
authors.lastname: Pintacuda
mesh_terms: D000150:Acinetobacter / Q000821:virology; D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D001435:Bacteriophages / Q000737:chemistry; D003460:Crystallization; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D019251:Nucleocapsid / Q000737:chemistry; D055503:Protein Multimerization; D011506:Proteins / Q000737:chemistry*; D011522:Protons
chemical_list: D000682:Amyloid; D011506:Proteins; D011522:Protons
keywords: magic-angle spinning; proton detection; resonance assignment; solid-state NMR spectroscopy