title: In-cell NMR characterization of the secondary structure populations of a disordered conformation of α-synuclein within E. coli cells.
abstract: α-Synuclein is a small protein strongly implicated in the pathogenesis of Parkinson's disease and related neurodegenerative disorders. We report here the use of in-cell NMR spectroscopy to observe directly the structure and dynamics of this protein within E. coli cells. To improve the accuracy in the measurement of backbone chemical shifts within crowded in-cell NMR spectra, we have developed a deconvolution method to reduce inhomogeneous line broadening within cellular samples. The resulting chemical shift values were then used to evaluate the distribution of secondary structure populations which, in the absence of stable tertiary contacts, are a most effective way to describe the conformational fluctuations of disordered proteins. The results indicate that, at least within the bacterial cytosol, α-synuclein populates a highly dynamic state that, despite the highly crowded environment, has the same characteristics as the disordered monomeric form observed in aqueous solution.
authors.affiliation: Institute of Structural and Molecular Biology, University College London and Birkbeck College, London, United Kingdom.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Waudby
authors.forename: Carlo
authors.initials: C
authors.lastname: Camilloni
authors.forename: Anthony W P
authors.initials: AW
authors.lastname: Fitzpatrick
authors.forename: Lisa D
authors.initials: LD
authors.lastname: Cabrita
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.forename: Michele
authors.initials: M
authors.lastname: Vendruscolo
authors.forename: John
authors.initials: J
authors.lastname: Christodoulou
mesh_terms: D004926:Escherichia coli / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017433:Protein Structure, Secondary; D051844:alpha-Synuclein / Q000737:chemistry*
chemical_list: D051844:alpha-Synuclein