title: Structural disorder of monomeric α-synuclein persists in mammalian cells.
abstract: Intracellular aggregation of the human amyloid protein α-synuclein is causally linked to Parkinson's disease. While the isolated protein is intrinsically disordered, its native structure in mammalian cells is not known. Here we use nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopy to derive atomic-resolution insights into the structure and dynamics of α-synuclein in different mammalian cell types. We show that the disordered nature of monomeric α-synuclein is stably preserved in non-neuronal and neuronal cells. Under physiological cell conditions, α-synuclein is amino-terminally acetylated and adopts conformations that are more compact than when in buffer, with residues of the aggregation-prone non-amyloid-β component (NAC) region shielded from exposure to the cytoplasm, which presumably counteracts spontaneous aggregation. These results establish that different types of crowded intracellular environments do not inherently promote α-synuclein oligomerization and, more generally, that intrinsic structural disorder is sustainable in mammalian cells.
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Francois-Xavier
authors.initials: FX
authors.lastname: Theillet
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Andres
authors.initials: A
authors.lastname: Binolfi
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Beata
authors.initials: B
authors.lastname: Bekei
authors.affiliation: Department of Chemical Physics, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Andrea
authors.initials: A
authors.lastname: Martorana
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Honor May
authors.initials: HM
authors.lastname: Rose
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Marchel
authors.initials: M
authors.lastname: Stuiver
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Silvia
authors.initials: S
authors.lastname: Verzini
authors.affiliation: Department of Molecular Physiology and Cell Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Dorothea
authors.initials: D
authors.lastname: Lorenz
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Marleen
authors.initials: M
authors.lastname: van Rossum
authors.affiliation: Department of Chemical Physics, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Daniella
authors.initials: D
authors.lastname: Goldfarb
authors.affiliation: In-Cell NMR Laboratory, Department of NMR-supported Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Robert-Rössle Strasse 10, 13125 Berlin, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Selenko
mesh_terms: D000107:Acetylation; D002460:Cell Line; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D006367:HeLa Cells; D006801:Humans; D042541:Intracellular Space / Q000737:chemistry* / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D051844:alpha-Synuclein / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein