title: Monitoring Conformational Landscape of Ovine Prion Protein Monomer Using Ion Mobility Coupled to Mass Spectrometry.
abstract: Prion protein is involved in deadly neurodegenerative diseases. Its pathogenicity is linked to its structural conversion (α-helix to β-strand transition). However, recent studies suggest that prion protein can follow a plurality of conversion pathways, which hints towards different conformers that might coexist in solution. To gain insights on the plasticity of the ovine prion protein (PrP) monomer, wild type (A136, R154, Q171), mutants and deletions of ARQ were studied by traveling wave ion mobility experiments coupled to mass spectrometry. In order to perform the analysis of a large body of data sets, we designed and evaluated the performance of a processing pipeline based on Driftscope peak detection and a homemade script for automated peak assignment, annotation, and quantification on specific multiply charged protein data. Using this approach, we showed that in the gas phase, PrPs are represented by at least three conformer families differing in both charge state distribution and collisional cross-section, in agreement with the work of Hilton et al. (2010). We also showed that this plasticity is borne both by the N- and C-terminal domains. Effect of protein concentration, pH and temperature were also assessed, showing that (1) pH does not affect conformer distributions, (2) protein concentration modifies the conformational landscape of one mutant (I208M) only, and (3) heating leads to other unfolded species and to a modification of the conformer intensity ratios. Graphical Abstract ᅟ.
authors.affiliation: Laboratoire de Chimie Physique, Université Paris Sud, CNRS UMR 8000, 91405, Orsay, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Van der Rest
authors.affiliation: Institut National de la Recherche Agronomique, UR892, Virologie Immunologie Moléculaires, Domaine de Vilvert, 78350, Jouy-en-Josas, France.
authors.forename: Human
authors.initials: H
authors.lastname: Rezaei
authors.affiliation: Laboratoire de Chimie Physique, Université Paris Sud, CNRS UMR 8000, 91405, Orsay, France. Frederic.Halgand@u-psud.fr.
authors.forename: Frédéric
authors.identifier: 0000-0002-5338-8235
authors.initials: F
authors.lastname: Halgand
mesh_terms: D000818:Animals; D002138:Calibration; D001330:Electronic Data Processing; D006863:Hydrogen-Ion Concentration; D000075663:Ion Mobility Spectrometry / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D009154:Mutation; D000072002:Prion Proteins / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D012756:Sheep; D013696:Temperature
chemical_list: D000072002:Prion Proteins
keywords: Dynamic; Ion mobility; Mass spectrometry; Ovine; Plasticity; Prion